HDTenEightyP/NetNews huggingface dataset:
From: broe@aardvark.ucs.uoknor.edu (Bruce Roe)
Subject: Re: Dissolution of DNA
News-Software: VAX/VMS VNEWS 1.41    
Organization: University of Oklahoma - University Computing Services
Distribution: bionet
Date: Tue, 14 Apr 1992 12:09:00 GMT

In article <12063.9204130857@subnode.susssys1.rdg.ac.uk>, skspoidn@susssys1.rdg.ac.uk (Mike Poidinger) writes...
>Help.
> 
>I am trying to redissolve DNA which has been glass-rod-extracted
>according to protocol 3 from Sambrook/Maniatis, and the %$#@ material
>wont dissolve, wont even come off the glass rod. I am trying to prepare
>medium sized genomic DNA from cell monolayers for southern Hybridization.
> 
>Any ideas on
>a) Getting the DNA off the glass or
>b) A better procedure for DNA preparation
> 
>would be greatly appreciated
> 
>Thanks in advance,
> 
>Mike Poidinger
>Dept of Microbiology
>University of Reading

Mike,
	Try removing the DNA from the glass rod with a sterile razor blade
and dissolving the DNA by gently rotating the DNA/buffer solution in at 4deg
overnight.  This worked many years ago for me with high mol.wt. bacterial
DNA.
	Bruce Roe	broe@aardvark.ucs.uoknor.edu
From: kowalski@sciborg.UWaterloo.ca (Paul Kowalski)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Biolistic expression of GUS with plant promoters
Keywords: GUS, biolystic, low-activity constitutive plant promoters
Date: 14 Apr 92 19:43:45 GMT
Distribution: bionet
Organization: University of Waterloo


  Has anyone in net-land have any experience in or knowledge of getting
expression of GUS in pBI101.1 with a low-activity constituative plant
promoter? I'm trying the APRT (apt) promoter from Arabidopsis and not
having much luck with biolistic-mediated transformation/transient
expression in tobacco suspension culture, Arabidopsis leaves and callus.
I'm using the usual DuPont system, and 1.6 micron gold particles, with
vectors containing GUS promoted by CaMV promoter as a positive control,
which works fine. 
Thanks in advance.....

Paul Kowalski
From: mcgraw%gandal.dnet@SERVER.UGA.EDU
Newsgroups: bionet.molbio.methds-reagnts
Subject: Thesit
Date: 9 Apr 92 14:26:23 GMT
Distribution: bionet


Hello Netters,

A recent short article in NAR (Ponce, M.R., and Nicol, J.L., 1992, Vol. 20, 
p.623) describes conditions for PCR amplification of long (>6 kb) DNA
fragments.  The 10X buffer contains 1% Thesit. The SIGMA catalog says
Thesit is a trade name for polyoxyethylene 9-lauryl ether.

Can anyone out there tell us more about Thesit in PCR?  If it's good, why
is it good?  Where did you get it?  Any comments appreciated.

				Thanks, 

				Al McGraw
				University of Georgia
Organization: University Hospital MEB, IMSDD, F.R. Germany
Date: Thursday, 16 Apr 1992 09:22:10 MEZ
From: Peter Hakenberg <NETADM@DBNMEB1.BITNET>
Newsgroups: bionet.molbio.methds-reagnts
Subject: how to add biotin to an oligo?

Does anyone have experience in adding biotin to a primer using the
ABI 392 Synthesizer. We want to use them for Sequencing with Dyna-beads
thx in advance
bye pi (Peter Hakenberg)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Breaking strepavidin/biotin bond.
From: yeager@sc2a.unige.ch (Mark Yeager)
Date: 16 Apr 92 10:42:47 +0200
Distribution: bionet,world
Organization: University of Geneva, Switzerland

In article <1992Apr13.133104.1@agri.huji.ac.il>, marder@agri.huji.ac.il
writes:

> I think that you may be in error here;  the main thing about iminobiotin is
> that it has a pK around 10 and only one form binds (strept)avidin, tho' I can't
> remember whether it is the protonated or unprotonated form.  In either case,

Must be the protonated form, if the binding is done around pH 7.

--   -.-
Mark Yeager  ->  yeager@cgeuge52.bitnet     yeager@sc2a.unige.ch
 University of Geneva, Dept. of Biochemistry, CH-1211 Geneva 4, Switzerland
From: suter@VAX.MPIZ-KOELN.MPG.DBP.DE
Newsgroups: bionet.molbio.methds-reagnts
Subject: tobacco sequence/a4t4 bx
Date: 15 Apr 92 17:18:06 GMT
Distribution: bionet


dear colleagues,

in the past few weeks i have cloned a piece of tobacco dna which has one
striking feature: it contains three repeated sequences within a stretch
of about 100 bp. I have compared this repeat with the databases: however,
nothing obvious came up. perhaps anybody out there could help ?
the repeat is:

	TACTTGTTGGCGTAAAATTTTGCA

especially this aaaatttt box looks so familiar (several co-workers suggested
this also) that I wondered whether any of the bells in your brains will also
start clanging away...
any suggestion welcome !

thanx ! clemens suter-crazzolara
From: yhsun%imb.dnet@LAS.AS.EDU.TW
Newsgroups: bionet.molbio.methds-reagnts
Subject: reuse of Dynabead-oligo(dT)?
Date: 12 Apr 92 04:15:01 GMT
Distribution: bionet

Can the oligo(dT)-Dynabead be reused? i.e. Can the oligo(dT)-dynabead
withstand NaOH wash? Is there a temperature limit for the elution step?

Henry Sun
From: byrne@soma.umdnj.edu (Bruce Byrne)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: RNase free DNase.
Date: 10 Apr 92 18:42:52 GMT

srckjel@chv.dsir.govt.nz writes:

>We are aiming to quantify RNA in an RNA prep. but having difficulty with
>co-isolated DNA.We want to use RNase free DNase to remove thisand would
>appreciate comments and advice on which companies provide the best product
>which is really (!) free of RNase.Thanks in advance.Jane Lancaster and Sonya
>Clark

   We used Promega's RQ1 with considerable success (NAR16:4165 1988)
You will want to titer your enzyme to your prep; any useful data will
be derived from lots of runs and preps with lots of DNA will require
substantial DNase.  A non-rt/PCR control demonstrates you've used enough
to kill DNA signal.

Bruce Byrne
Newsgroups: bionet.molbio.methds-reagnts
From: txt15@po.CWRU.Edu (Tao Tao)
Subject: Re: DIGESTION OF PCR PRODUCTS
Organization: Case Western Reserve University, Cleveland, Ohio, (USA)
Date: Sat, 11 Apr 92 18:04:38 GMT


I don't know what method has been used in purifying the PCR products after
the Proteinase, any phenol extraction? what about the residual phenol?

Another question is the bases used in the end of the primers, there should be
2-3 bp of G and/or C.  Also the digest temperature might also have some effect.

These are the tricky steps I could think about.

My suggestion is: Proteinase treatment;
		  phenol extraction;
		  Spun-column purification;
		  restriction digestion.

Hope this helps.

Tao
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Breaking strepavidin/biotin bond.
From: marder@agri.huji.ac.il
Date: 13 Apr 92 13:31:04 IST
Distribution: bionet,world

In article <9204121706.AA25167@genbank.bio.net>, MIBJOEL@UKCC.UKY.EDU (Joel Guthridge) writes:
> Tom Schneider recently asked how one could break the strepavidin/biotin
> interaction to recover the biotin conjugated material.  If I remember
> correctly, one can use iminobiotin instead of biotin.  The advantage
> is that the iminobiotin/strepavidin interaction has a much lower affinity
> than the biotin/strepavidin interaction...  allowing less harsh elution
> methods to be employed.  I think Sigma and Pierce (as well as others I'm
> sure) sell NHS- or hydrazine based iminobiotin for conjugate preparation.

I think that you may be in error here;  the main thing about iminobiotin is
that it has a pK around 10 and only one form binds (strept)avidin, tho' I can't
remember whether it is the protonated or unprotonated form.  In either case,
you have to expose your sample to fairly alkaline conditions for the
binding or unbinding stage - not too bad for most proteins, but perhaps
problematic for nucleic acids.  Iminobiotin and derivitive labelling kits
are listed in the Sigma catalogue and they give the following refs:-
	Heney G et al. (1981) Analytical Biochem. 114,92.
	Orr GA et al. (1981) JBC 256,761.
Another biotin derivative listed in the Sigma catalogue is Diaminobiotin
(Ref. Green NM; 1966, Biochem. J., 101, 774).  Perhaps it works in a
similar way.
--
                                 '      Jonathan B. Marder
Internet: MARDER@AGRI.HUJI.AC.IL       Department of Agricultural Botany
Bitnet:   MARDER@HUJIAGRI         /\/  The Hebrew University of Jerusalem
Phone:    (08 or +9728) 481918   /  \  Faculty of Agriculture
Fax:      (08 or +9728) 467763   /      P.O.Box 12, Rehovot 76100, ISRAEL
From: dak@biosys.apldbio.COM (David Knorr)
Newsgroups: bionet.molbio.methds-reagnts
Subject: breaking streptavidin/biotin linkage
Date: 13 Apr 92 17:51:54 GMT
Organization: Applied Biosystems, Inc., Foster City, CA


 	 Tom Schneider wonders about breaking the biotin/streptavidin
bond.  Hot ( ~95 C ) dimethyformamide works well.  Just boiling in
water doesnt seem to work as well.

***********************
* Dave Knorr          *
* DAK@apldbio.com     *    
* voice: 415 570 6667 *
***********************
From: skspoidn@susssys1.rdg.ac.uk (Mike Poidinger)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Pcr sequencing
Date: 8 Apr 92 15:10:05 GMT
Distribution: bionet

(coming late to the conversation, in response to <9204071426.AA04494 7/4/92>)
I have used, and got clean sequence results with, the Promega
PCR sequencing kit, which utilizes modified Taq polymerase. I have used
5' end labelling with 32P of PCR primers and internal sequencing primers.
PCR product (800bp from genomic DNA) was milk-glass cleaned, and small
 quantities (50 fmol) were
sequenced according to product specifications. Sequence is (sort of :-) readable
 to
within a few bases of the primer.

I have found that the cycling protocol depends on the thermal cycler being used,
especially wrt whether it measures plate or tube temperatures. Setting the
temp control to tube can mean long wait periods at plateau temperatures before
the cycle countdown begins.

 (I THINK:) If a low (<55 deg) annealing temp is used, Considerable
strand elongation and termination can occur during this temp stabilization
 period,
and at this temp, the system is sensitive to secondary structure false
 termination,
resulting in strong cross banding, poor sequence etc. Taq can incorporate 90
 bases/min
at 37 deg

Idealy, annealing temp should be a few degrees below melting temp, and should
 only
be held for 1 minute at the most. I use 30 secs.

Promega kit, including ddNTP's, PNK, Taq pol for 100 reactions costs 117 pounds
 stirling
From: kychen@chem.nthu.edu.tw (Kuang-Yu Chen (035)715131-3352)
Newsgroups: bionet.molbio.evolution
Subject: help!!!
Date: 7 Apr 92 02:30:11 GMT
Organization: National Tsing Hua University (HsinChu)

Dear Mr. & Mrs. :

I am applying for admission to Marine-Estuarine-Environmental
Science Graduate Program at the Univ. of Maryland. The Graduate
Admissions Committee informed me that it is not possible for me
to be admitted without a major advisor.

I want to contact some faculty members in MEES Program, but I
have no their email addr.. It's urgent for me.

The list of the professors is in the following :
(1) Dr. Bashore, Terry,   NATS/UMES
(2) Dr. Hoogland, John L.,  AEL/CEES
(3) Dr. Inouye, David W., ZOOL/UMCP
(4) Dr. Kuenzel, Wayne J., POUL/UMCP
(5) Dr. Mench, Joy A., POUL/UMCP
(6) Dr. Provine, Robert, PSYC/UMBC
(7) Dr. Rohwer, Frank C., AEL/CEES
(8) Dr. Costanza, Robert, CBL/CEES

If someone knows their email addr., please tell me or you can
give me other advices.

Thanks in advance.

______________________
kychen@y5.nthu.edu.tw
886-35-715131 ext3352
Room 265, Dept. of Chemistry, National Tsing Hua U., 
Hsinchu, Taiwan 300
From: philstas@VUB.AC.BE (Stas Philippe)
Newsgroups: bionet.molbio.hiv
Subject: Last try
Date: 16 Apr 92 08:51:53 GMT
Article-I.D.: genbank.9204161035.AA18914
Distribution: bionet


Hy,

Let's say this is the last time i'm gonna try
to submit a question.

Anyone knows references about the effect of the V3 loop on infectivity of
HIV-1. I found one article (Wahlberg et al., AIDS research and human
retrovirusses Vol.7, 983) were different cases of HIV infection
without variation in de V3 loop showed different infection and
occurance of the disease. Other patients showed variation of their
own V3 loop but it didn'taffect the occuranced of their disease.

Could anybody sendme more information about this? (if there is
any)

-------------------------------------------------------------
-  Philippe Stas                        philstas@vub.ac.be  -
-  Dept. of Cellular Immunology                              -
-  Free University of Brussels          tel: 32-2-359.03.58 -
-  Paardenstraat 65                     Fax: 32-2-359.03.90 -
-  B1640 St.Genesius Rode                                   -
-------------------------------------------------------------
From: philstas@VUB.AC.BE (Stas Philippe)
Newsgroups: bionet.molbio.hiv
Subject: HELP
Date: 14 Apr 92 16:32:28 GMT
Article-I.D.: genbank.9204141701.AA17447
Distribution: bionet


Can anybody tell me why I don't get any mail at all from this list?
Can someone do something against it.

Thanks

Johor

"If Man is five, Then the devil is six"
"And if the devil is six, Then GOD is seven"
"This monkey is going to heaven"
-------------------------------------------------------------
-  Philippe Stas                        philstas@vub.ac.be  -
-  Dept. of Cellular Immunology                              -
-  Free University of Brussels          tel: 32-2-359.03.58 -
-  Paardenstraat 65                     Fax: 32-2-359.03.90 -
-  B1640 St.Genesius Rode                                   -
-------------------------------------------------------------
From: philstas@VUB.AC.BE (Stas Philippe)
Newsgroups: bionet.molbio.hiv
Subject: anybody out there?
Date: 13 Apr 92 13:55:47 GMT
Distribution: bionet


I subscribed to this list one week ago.
Untill now I didn't rreceive any mail and posted two
mails myself.

On the review-list I dopn't seem to find any users
Is there anybody else than me on this list?
How can I get the complete list of subscribed users?

Thanks for letting me know, via the list and at my adress.

-------------------------------------------------------------
-  Philippe Stas                        philstas@vub.ac.be  -
-  Dept. of Cellular Immunology                              -
-  Free University of Brussels          tel: 32-2-359.03.58 -
-  Paardenstraat 65                     Fax: 32-2-359.03.90 -
-  B1640 St.Genesius Rode                                   -
-------------------------------------------------------------
From: VICE@PASTEUR.BITNET (Mike Robertson)
Newsgroups: bionet.molbio.hiv
Subject: pSVArt sequence
Date: 9 Apr 92 09:06:48 GMT
Article-I.D.: genbank.9204090906.AA23528
Distribution: bionet


Does anyone have the full sequence of pSVart (Rosen, et al, PNAS 85:2071-
2075, 1988)?  I have pieced it together from pSV2 sequences and the published
sequence of the HIV-1 rev sequence from the AIDS Database, but I don't know
precisely where the 5' end of the cDNA insert is or if there are any other
hidden surprises.
Thanks.
From: MA11@PHX.CAM.AC.UK
Newsgroups: bionet.molbio.genbank
Subject: (none)
Date: 17 Apr 92 12:04:40 GMT
Article-I.D.: UK.A58E856B48B28440
Distribution: bionet

I wish to take up the comment made in Robison's bulletin about the need
for meta-entries in the databanks. I could not agree more.
It is common place for the same or different labs to sequence the "same"
gene from the same species and for these to be represented by individual
AC's in the databases. So they should be. I am strongly of the opinion
that such entries should never be merged, each AC should represent an
"unit" of experimental data. These different sequences of the "same" gene
may differ in many ways - they may well not be coterminous, one may extend
further 5' or 3' than the other; they may well differ in the regions
common to them, these differences may be due to allelic differences or
to experimental error; the sequences may not necessarily be represented
in the data bank as the same strand (this is especially likely where the
overlap between two AC's is because two labs have sequenced adjacent genes
transcribed in opposite directions).
For the majority of users what is needed is  one sequence showing the maximum
amount of information concerning a particular gene. To construct this from
the databanks would now be a lot of work. For example a search on the two
terms 'DROSOPHILA MELANOGASTER' AND 'ALCOHOL DEHYDROGENASE' will return
35 different AC's from the databanks. Unless one has specialist knowledge about
this gene unravelling these is a nightmare. It would be a grave mistake
to attempt to make a single virtual sequence of the fly ADH (a mistake,
indeed, that I have made, see AC Z00030) for every time a new sequence
is made avaliable (or an old one corrected) this virtual sequence must be
changed.
Instead, what I want is a syntax in which I can describe this virtual sequence
in terms of the 35 different accessions and a program that will
take this description (and the databank) and build me the sequence whilst
I drink the (next) cup of coffee. This sequence will be fully annotated -
conflicts between different primary entries will be noted, as will be
sequence features.
I am not sure what is the "meta-sequence" in Robison's sense - the
description or the sequence constructed with this description - but I am
sure that these tools are needed. I would see it as a function of
specialized curators or genetic database curators to write and maintain
the descriptions of the virtual sequence. However before they (we)
can do so they (we) need (a) the syntax for the descriptions, (b) the
programs to read the descriptions and output the annotated virtual
sequences and (c) a workbench for writing/maintaining the descriptions.
Michael Ashburner, Department of Genetics, Cambridge, England.
ma11@cam.phx.ac.uk.
From: GenBank-Updates@genbank.bio.net
Newsgroups: bionet.molbio.genbank.updates
Subject: Human endothelial leukocyte adhesion molecule I (ELAM1) gene, exon
Date: 10 Apr 92 10:05:41 GMT
Distribution: bionet
Checksum: 56808     2

LOCUS       HUMELAM4      400 bp ds-DNA             PRI       10-APR-1992
DEFINITION  Human endothelial leukocyte adhesion molecule I (ELAM1) gene, exon
            4.
ACCESSION   M61889 M58017
KEYWORDS    endothelial leukocyte adhesion molecule I.
SEGMENT     4 of 9
SOURCE      Homo sapiens DNA.
  ORGANISM  Homo sapiens
            Eukaryota; Animalia; Chordata; Vertebrata; Mammalia; Theria;
            Eutheria; Primates; Haplorhini; Catarrhini; Hominidae.
REFERENCE   1  (bases 1 to 400)
  AUTHORS   Collins,T., Williams,A., Johnston,G.I., Kim,J., Eddy,R., Shows,T.,
            Gimbrone,M.A. and Bevilacqua,M.P.
  TITLE     Structure and chromosomal location of the gene for
            endothelial-leukocyte adhesion molecule 1
  JOURNAL   J. Biol. Chem. 266, 2466-2473 (1991)
  STANDARD  full automatic
COMMENT     From EMBL    entry HSELAM4;  dated 07-MAR-1991.
FEATURES             Location/Qualifiers
     exon            140..325
                     /number=4
                     /gene="ELAM1"
BASE COUNT      112 a     84 c     78 g    126 t
ORIGIN
        1 agtcaatttt acaaattcca ttagaaattt tataaatttt taaactaaaa aaaaaaaaac
       61 caaacgtagt tctagttttc agtgttctta tctgttaaag gtgagagatg agcagaactg
      121 atttgggttt ctttttcagt tgtgaactgt acagccctgg aatcccctga gcatggaagc
      181 ctggtttgca gtcacccact gggaaacttc agctacaatt cttcctgctc tatcagctgt
      241 gataggggtt acctgccaag cagcatggag accatgcagt gtatgtcctc tggagaatgg
      301 agtgctccta ttccagcctg caatggtaaa tccctctgag aatgcacaga acgttctgta
      361 actcatccct cattgcttct gttttttgat tcaacttaca
//
From: GenBank-Updates@genbank.bio.net
Newsgroups: bionet.molbio.genbank.updates
Subject: Murine coronavirus RNA, 5' end.
Date: 10 Apr 92 10:07:58 GMT
Distribution: bionet
Checksum: 22871     1

LOCUS       MHVGENRNA     150 bp ss-RNA             VRL       10-APR-1992
DEFINITION  Murine coronavirus RNA, 5' end.
ACCESSION   M27460
KEYWORDS    .
SOURCE      Murine hepatitis virus (strain JHM) RNA.
  ORGANISM  Murine hepatitis virus
            Viridae; ss-RNA enveloped viruses; Positive strand RNA virus;
            Coronaviridae.
REFERENCE   1  (bases 1 to 150)
  AUTHORS   Soe,L.H., Shieh,C.-K., Makino,S., Chang,M.-F., Stohlman,S.A. and
            Lai,M.M.
  TITLE     Murine coronavirus 5'-end genomic RNA sequence reveals mechanism of
            leader-primed transcription
  JOURNAL   Adv. Exp. Med. Biol. 218, 73-81 (1987)
  STANDARD  full staff_entry
BASE COUNT       33 a     36 c     29 g     52 t
ORIGIN
        1 tataagagtg attggcgtcc gtacgtaccc tctctactct aaaactcttg tagtttaaat
       61 ctaatctaat ctaaacttta taaacggcac ttcctgcgtg tccatgcccg tgggcctggt
      121 cttgtcatag tgctgacatt tgtggttcct
//
From: GenBank-Updates@genbank.bio.net
Newsgroups: bionet.molbio.genbank.updates
Subject: Treponema sp. small subunit ribosomal RNA sequence.
Date: 10 Apr 92 10:14:19 GMT
Distribution: bionet
Checksum: 63867     3

LOCUS       TRPSSRNA     1363 bp ss-rRNA            RNA       10-APR-1992
DEFINITION  Treponema sp. small subunit ribosomal RNA sequence.
ACCESSION   M59179
KEYWORDS    small subunit ribosomal RNA.
SOURCE      Treponema sp. (library: ATCC 43811) rRNA.
  ORGANISM  Treponema sp.
            Prokaryota; Bacteria; Gracilicutes; Scotobacteria; Spirochetes;
            Spirochaetales; Spirochaetaceae.
REFERENCE   1  (bases 1 to 1363)
  AUTHORS   Paster,B.J., Dewhirst,F.E., Weisburg,W.G., Tordoff,L.A.,
            Fraser,G.J., Hespell,R.B., Stanton,T.B., Zablen,L.B., Mandelco,L.
            and Woese,C.R.
  TITLE     Phylogenetic analysis of the spirochetes
  JOURNAL   J. Bacteriol. 173, 6101-6109 (1991)
  STANDARD  simple staff_entry
BASE COUNT      361 a    279 c    418 g    275 t     30 others
ORIGIN
        1 ttaataatgg agggtttgat tctggctcag agcgaacgct ggcagcgngt tttaggcatg
       61 caagtcgaac ggcagcggga tagagcttgc tctattgccg gcgagtggca gacgggtgag
      121 taacgcgtag gaatctgccn ataggactgg gatagccaag agaaattttg ggtaataccn
      181 gatgatatcg agagatcaaa ggggcatttg ccntacctgt ggatgagcct acgtccnatc
      241 agctagttgg ttaggtaaag gctnaccaag gctaagacgg gtagccggct tgagagagtg
      301 gccggccaca agggaactga gacacggtcc nnactcctac gggaggcagc agccaagaat
      361 attccgcaat gggggaaacc cnnacggagc gacgcngcgt gaaggaagaa ggcattcgtg
      421 ttgtaaactn cttttctaaa ggaagaataa gtgtgggagg aaatgccnac atgatgacgg
      481 tactttagga ataagccccg gctaattacg tgccagcagc cncggtaata cgtaagggnn
      541 nagcgttgct cggaattact gggcgtaaag ggcgtgtagg cngtttatta ggccgagtgt
      601 gaaagaccgg nnnncnactt cggnaacgcg ctcgnnaccg atagactaga atctagggga
      661 aggtagtgga atttcctgtg tagcggtgga atgcatagat atgggaagga acaccaaagg
      721 cgaaggcagc tacctatgct aagattgacg ctgaggcgcg aaagtgtggg gatcaaatag
      781 gattagatac cctagtagtc cacactgtaa acgatgtaca cttagagttg cgcgagagcg
      841 tngttctgca gcaaacgcgt taagtgtacc gcctgaggag tacggccgca aggctgaaac
      901 tcaaagaaat tgacgggggc cngcacaagc ggtggagcac gtggattaat tcgatgntaa
      961 gcgaagaacc ttaccaaggc ttgacatata ctggaatgtt gtggaaacat ggctagcctt
     1021 cgggccagta tacaggtgct gcatggctgt cgtcagctcg tgtcgtgaga tgttgggtta
     1081 agtcccncaa cgagcgcaac ccttgttttc agttgctact attaagttag aggactctgg
     1141 aaagactgcc ggcgacaagt cgggggaagg aggggacgac gtcaagtcat catggccctt
     1201 atgtcttggg cttcacacat gctacaatgg tgcggacagc gggaagcgaa gtcgcgaggc
     1261 ggagctaaac ctccaaaccg tacctcaata cggattgcac tctgaaactc gagtgcatga
     1321 agccggaatc gctagtaatc gcgggtcagc aataccgcgg tga
//