pdCASclos

LOCUS      pdCASclos              9056 bp ds-DNA     

circular SYN 12-MAY-2021
DEFINITION  Transcriptional repression for gene Spo0A 

(CAC-2011) in Clostridium 
            acetobutylicum ATCC 824.
ACCESSION   .
VERSION     .
KEYWORDS    pdCASclos
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9056)
  AUTHORS   Li Q, Chen J, Minton NP, Zhang Y, Wen Z, Liu 

J, Yang H, Zeng Z, Ren 
            X, Yang J, Gu Y, Jiang W, Jiang Y, Yang S
  TITLE     CRISPR-based genome editing and expression 

control systems in 
            Clostridium acetobutylicum and Clostridium 

beijerinckii.
  JOURNAL   Biotechnol J. 2016 May 23. doi: 

10.1002/biot.201600053.
  PUBMED    27213844
REFERENCE   2  (bases 1 to 9056)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 

1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9056
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        14..48
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry 

of Standard Biological 
                     Parts BBa_J23119) modified to end 

with an SpeI site"
     misc_RNA        69..144
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the 

Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             281..4387
                     /codon_start=1
                     /product="catalytically dead mutant 

of the Cas9 
                     endonuclease from the Streptococcus 

pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding 

protein that lacks 
                     endonuclease activity due to the 

D10A mutation in the RuvC 
                     catalytic domain and the H840A 

mutation in the HNH 
                     catalytic domain"
                     

/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSI

KK
                     

NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE

ES
                     

FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHM

IK
                     

FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSR

RL
                     

ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLL

AQ
                     

IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKAL

VR
                     

QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNRED

LL
                     

RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLA

RG
                     

NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLY

EY
                     

FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIEC

FD
                     

SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIE

ER
                     

LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRN

FM
                     

QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVM

GR
                     

HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEK

LY
                     

LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDN

VP
                     

SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQIT

KH
                     

VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDA

YL
                     

NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFF

KT
                     

EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGF

SK
                     

ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELL

GI
                     

TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKG

NE
                     

LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVIL

AD
                     

ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE

VL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(4397..4414)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called 

M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4397..4413)
                     /label=M13 fwd
                     /note="common sequencing primer, one 

of multiple similar 
                     variants"
     primer_bind     complement(4406..4428)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4622..4641)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence 

yeast selectable 
                     marker"
     primer_bind     4741..4763
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4801..4819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of 

EcoRI site, forward 
                     primer"
     promoter        4887..4991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4992..5852
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to 

ampicillin, carbenicillin, and
                     related antibiotics"
                     

/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVG

YI
                     

ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE

YS
                     

PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLD

RW
                     

EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLR

SA
                     

LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIG

AS
                     LIKHW"
     primer_bind     complement(5210..5229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, 

reverse primer"
     rep_origin      6023..6611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number 

ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6512..6531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward 

primer"
     primer_bind     6765..6782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6899..6920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite 

activator protein"
                     /note="CAP binding activates 

transcription in the presence 
                     of cAMP."
     promoter        6935..6965
                     /label=lac promoter
                     /note="promoter for the E. coli lac 

operon"
     protein_bind    6973..6989
                     /label=lac operator
                     /bound_moiety="lac repressor encoded 

by lacI"
                     /note="The lac repressor binds to 

the lac operator to 
                     inhibit transcription in E. coli. 

This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-

thiogalactopyranoside (IPTG)."
     primer_bind     6978..7000
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6997..7013
                     /label=M13 rev
                     /note="common sequencing primer, one 

of multiple similar 
                     variants"
     primer_bind     6997..7013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called 

M13-rev"
ORIGIN
        1 agcttggctg cagttgacag ctagctcagt cctaggtata 

atactagttt taattgcaga
       61 tgataatagt tttagagcta gaaatagcaa gttaaaataa 

ggctagtccg ttatcaactt
      121 gaaaaagtgg caccgagtcg gtgctttttt gcggccgcta 

taaaatataa ataattttct
      181 aaaaaactta acttcatgtg aaaagtttgt taaaatataa 

atgagcacgt taatcattta
      241 acatagataa ttaaatagta aaagggagtg tcgactcgag 

atggataaga aatactcaat
      301 aggcttagct atcggcacaa atagcgtcgg atgggcggtg 

atcactgatg aatataaggt
      361 tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc 

cacagtatca aaaaaaatct
      421 tataggggct cttttatttg acagtggaga gacagcggaa 

gcgactcgtc tcaaacggac
      481 agctcgtaga aggtatacac gtcggaagaa tcgtatttgt 

tatctacagg agattttttc
      541 aaatgagatg gcgaaagtag atgatagttt ctttcatcga 

cttgaagagt cttttttggt
      601 ggaagaagac aagaagcatg aacgtcatcc tatttttgga 

aatatagtag atgaagttgc
      661 ttatcatgag aaatatccaa ctatctatca tctgcgaaaa 

aaattggtag attctactga
      721 taaagcggat ttgcgcttaa tctatttggc cttagcgcat 

atgattaagt ttcgtggtca
      781 ttttttgatt gagggagatt taaatcctga taatagtgat 

gtggacaaac tatttatcca
      841 gttggtacaa acctacaatc aattatttga agaaaaccct 

attaacgcaa gtggagtaga
      901 tgctaaagcg attctttctg cacgattgag taaatcaaga 

cgattagaaa atctcattgc
      961 tcagctcccc ggtgagaaga aaaatggctt atttgggaat 

ctcattgctt tgtcattggg
     1021 tttgacccct aattttaaat caaattttga tttggcagaa 

gatgctaaat tacagctttc
     1081 aaaagatact tacgatgatg atttagataa tttattggcg 

caaattggag atcaatatgc
     1141 tgatttgttt ttggcagcta agaatttatc agatgctatt 

ttactttcag atatcctaag
     1201 agtaaatact gaaataacta aggctcccct atcagcttca 

atgattaaac gctacgatga
     1261 acatcatcaa gacttgactc ttttaaaagc tttagttcga 

caacaacttc cagaaaagta
     1321 taaagaaatc ttttttgatc aatcaaaaaa cggatatgca 

ggttatattg atgggggagc
     1381 tagccaagaa gaattttata aatttatcaa accaatttta 

gaaaaaatgg atggtactga
     1441 ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc 

aagcaacgga cctttgacaa
     1501 cggctctatt ccccatcaaa ttcacttggg tgagctgcat 

gctattttga gaagacaaga
     1561 agacttttat ccatttttaa aagacaatcg tgagaagatt 

gaaaaaatct tgacttttcg
     1621 aattccttat tatgttggtc cattggcgcg tggcaatagt 

cgttttgcat ggatgactcg
     1681 gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa 

gttgtcgata aaggtgcttc
     1741 agctcaatca tttattgaac gcatgacaaa ctttgataaa 

aatcttccaa atgaaaaagt
     1801 actaccaaaa catagtttgc tttatgagta ttttacggtt 

tataacgaat tgacaaaggt
     1861 caaatatgtt actgaaggaa tgcgaaaacc agcatttctt 

tcaggtgaac agaagaaagc
     1921 cattgttgat ttactcttca aaacaaatcg aaaagtaacc 

gttaagcaat taaaagaaga
     1981 ttatttcaaa aaaatagaat gttttgatag tgttgaaatt 

tcaggagttg aagatagatt
     2041 taatgcttca ttaggtacct accatgattt gctaaaaatt 

attaaagata aagatttttt
     2101 ggataatgaa gaaaatgaag atatcttaga ggatattgtt 

ttaacattga ccttatttga
     2161 agatagggag atgattgagg aaagacttaa aacatatgct 

cacctctttg atgataaggt
     2221 gatgaaacag cttaaacgtc gccgttatac tggttgggga 

cgtttgtctc gaaaattgat
     2281 taatggtatt agggataagc aatctggcaa aacaatatta 

gattttttga aatcagatgg
     2341 ttttgccaat cgcaatttta tgcagctgat ccatgatgat 

agtttgacat ttaaagaaga
     2401 cattcaaaaa gcacaagtgt ctggacaagg cgatagttta 

catgaacata ttgcaaattt
     2461 agctggtagc cctgctatta aaaaaggtat tttacagact 

gtaaaagttg ttgatgaatt
     2521 ggtcaaagta atggggcggc ataagccaga aaatatcgtt 

attgaaatgg cacgtgaaaa
     2581 tcagacaact caaaagggcc agaaaaattc gcgagagcgt 

atgaaacgaa tcgaagaagg
     2641 tatcaaagaa ttaggaagtc agattcttaa agagcatcct 

gttgaaaata ctcaattgca
     2701 aaatgaaaag ctctatctct attatctcca aaatggaaga 

gacatgtatg tggaccaaga
     2761 attagatatt aatcgtttaa gtgattatga tgtcgatgca 

attgttccac aaagtttcct
     2821 taaagacgat tcaatagaca ataaggtctt aacgcgttct 

gataaaaatc gtggtaaatc
     2881 ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa 

aactattgga gacaacttct
     2941 aaacgccaag ttaatcactc aacgtaagtt tgataattta 

acgaaagctg aacgtggagg
     3001 tttgagtgaa cttgataaag ctggttttat caaacgccaa 

ttggttgaaa ctcgccaaat
     3061 cactaagcat gtggcacaaa ttttggatag tcgcatgaat 

actaaatacg atgaaaatga
     3121 taaacttatt cgagaggtta aagtgattac cttaaaatct 

aaattagttt ctgacttccg
     3181 aaaagatttc caattctata aagtacgtga gattaacaat 

taccatcatg cccatgatgc
     3241 gtatctaaat gccgtcgttg gaactgcttt gattaagaaa 

tatccaaaac ttgaatcgga
     3301 gtttgtctat ggtgattata aagtttatga tgttcgtaaa 

atgattgcta agtctgagca
     3361 agaaataggc aaagcaaccg caaaatattt cttttactct 

aatatcatga acttcttcaa
     3421 aacagaaatt acacttgcaa atggagagat tcgcaaacgc 

cctctaatcg aaactaatgg
     3481 ggaaactgga gaaattgtct gggataaagg gcgagatttt 

gccacagtgc gcaaagtatt
     3541 gtccatgccc caagtcaata ttgtcaagaa aacagaagta 

cagacaggcg gattctccaa
     3601 ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt 

gctcgtaaaa aagactggga
     3661 tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct 

tattcagtcc tagtggttgc
     3721 taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt 

aaagagttac tagggatcac
     3781 aattatggaa agaagttcct ttgaaaaaaa tccgattgac 

tttttagaag ctaaaggata
     3841 taaggaagtt aaaaaagact taatcattaa actacctaaa 

tatagtcttt ttgagttaga
     3901 aaacggtcgt aaacggatgc tggctagtgc cggagaatta 

caaaaaggaa atgagctggc
     3961 tctgccaagc aaatatgtga attttttata tttagctagt 

cattatgaaa agttgaaggg
     4021 tagtccagaa gataacgaac aaaaacaatt gtttgtggag 

cagcataagc attatttaga
     4081 tgagattatt gagcaaatca gtgaattttc taagcgtgtt 

attttagcag atgccaattt
     4141 agataaagtt cttagtgcat ataacaaaca tagagacaaa 

ccaatacgtg aacaagcaga
     4201 aaatattatt catttattta cgttgacgaa tcttggagct 

cccgctgctt ttaaatattt
     4261 tgatacaaca attgatcgta aacgatatac gtctacaaaa 

gaagttttag atgccactct
     4321 tatccatcaa tccatcactg gtctttatga aacacgcatt 

gatttgagtc agctaggagg
     4381 tgactgaccc gggggaactg gccgtcgttt tacaacgtcg 

tgactgggaa aaccctggcg
     4441 ttacccaact taatcgcctt gcagcacatc cccctttcgc 

cagctggcgt aatagcgaag
     4501 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 

gaatggcgaa tggcgcctga
     4561 tgcggtattt tctccttacg catctgtgcg gtatttcaca 

ccgcatatgg tgcactctca
     4621 gtacaatctg ctctgatgcc gcatagttaa gccagccccg 

acacccgcca acacccgctg
     4681 acgcgccctg acgggcttgt ctgctcccgg catccgctta 

cagacaagct gtgaccgtct
     4741 ccgggagctg catgtgtcag aggttttcac cgtcatcacc 

gaaacgcgcg agacgaaagg
     4801 gcctcgtgat acgcctattt ttataggtta atgtcatgat 

aataatggtt tcttagacgt
     4861 caggtggcac ttttcgggga aatgtgcgcg gaacccctat 

ttgtttattt ttctaaatac
     4921 attcaaatat gtatccgctc atgagacaat aaccctgata 

aatgcttcaa taatattgaa
     4981 aaaggaagag tatgagtatt caacatttcc gtgtcgccct 

tattcccttt tttgcggcat
     5041 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa 

agtaaaagat gctgaagatc
     5101 agttgggtgc acgagtgggt tacatcgaac tggatctcaa 

cagcggtaag atccttgaga
     5161 gttttcgccc cgaagaacgt tttccaatga tgagcacttt 

taaagttctg ctatgtggcg
     5221 cggtattatc ccgtattgac gccgggcaag agcaactcgg 

tcgccgcata cactattctc
     5281 agaatgactt ggttgagtac tcaccagtca cagaaaagca 

tcttacggat ggcatgacag
     5341 taagagaatt atgcagtgct gccataacca tgagtgataa 

cactgcggcc aacttacttc
     5401 tgacaacgat cggaggaccg aaggagctaa ccgctttttt 

gcacaacatg ggggatcatg
     5461 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc 

cataccaaac gacgagcgtg
     5521 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa 

actattaact ggcgaactac
     5581 ttactctagc ttcccggcaa caattaatag actggatgga 

ggcggataaa gttgcaggac
     5641 cacttctgcg ctcggccctt ccggctggct ggtttattgc 

tgataaatct ggagccggtg
     5701 agcgtgggtc tcgcggtatc attgcagcac tggggccaga 

tggtaagccc tcccgtatcg
     5761 tagttatcta cacgacgggg agtcaggcaa ctatggatga 

acgaaataga cagatcgctg
     5821 agataggtgc ctcactgatt aagcattggt aactgtcaga 

ccaagtttac tcatatatac
     5881 tttagattga tttaaaactt catttttaat ttaaaaggat 

ctaggtgaag atcctttttg
     5941 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 

ccactgagcg tcagaccccg
     6001 tagaaaagat caaaggatct tcttgagatc ctttttttct 

gcgcgtaatc tgctgcttgc
     6061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 

ggatcaagag ctaccaactc
     6121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc 

aaatactgtc cttctagtgt
     6181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc 

gcctacatac ctcgctctgc
     6241 taatcctgtt accagtggct gctgccagtg gcgataagtc 

gtgtcttacc gggttggact
     6301 caagacgata gttaccggat aaggcgcagc ggtcgggctg 

aacggggggt tcgtgcacac
     6361 agcccagctt ggagcgaacg acctacaccg aactgagata 

cctacagcgt gagctatgag
     6421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta 

tccggtaagc ggcagggtcg
     6481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 

ctggtatctt tatagtcctg
     6541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 

atgctcgtca ggggggcgga
     6601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt 

cctggccttt tgctggcctt
     6661 ttgctcacat gttctttcct gcgttatccc ctgattctgt 

ggataaccgt attaccgcct
     6721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 

gcgcagcgag tcagtgagcg
     6781 aggaagcgga agagcgccca atacgcaaac cgcctctccc 

cgcgcgttgg ccgattcatt
     6841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg 

cagtgagcgc aacgcaatta
     6901 atgtgagtta gctcactcat taggcacccc aggctttaca 

ctttatgctt ccggctcgta
     6961 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg 

aaacagctat gaccatgatt
     7021 acgccaagct ttggctaaca cacacgccat tccaaccaat 

agttttctcg gcataaagcc
     7081 atgctctgac gcttaaatgc actaatgcct taaaaaaaca 

ttaaagtcta acacactaga
     7141 cttatttact tcgtaattaa gtcgttaaac cgtgtgctct 

acgaccaaaa gtataaaacc
     7201 tttaagaact ttcttttttc ttgtaaaaaa agaaactaga 

taaatctctc atatctttta
     7261 ttcaataatc gcatcagatt gcagtataaa tttaacgatc 

actcatcatg ttcatattta
     7321 tcagagctcg tgctataatt atactaattt tataaggagg 

aaaaaataaa gagggttata
     7381 atgaacgaga aaaatataaa acacagtcaa aactttatta 

cttcaaaaca taatatagat
     7441 aaaataatga caaatataag attaaatgaa catgataata 

tctttgaaat cggctcagga
     7501 aaagggcatt ttacccttga attagtacag aggtgtaatt 

tcgtaactgc cattgaaata
     7561 gaccataaat tatgcaaaac tacagaaaat aaacttgttg 

atcacgataa tttccaagtt
     7621 ttaaacaagg atatattgca gtttaaattt cctaaaaacc 

aatcctataa aatatttggt
     7681 aatatacctt ataacataag tacggatata atacgcaaaa 

ttgtttttga tagtatagct
     7741 gatgagattt atttaatcgt ggaatacggg tttgctaaaa 

gattattaaa tacaaaacgc
     7801 tcattggcat tatttttaat ggcagaagtt gatatttcta 

tattaagtat ggttccaaga
     7861 gaatattttc atcctaaacc taaagtgaat agctcactta 

tcagattaaa tagaaaaaaa
     7921 tcaagaatat cacacaaaga taaacagaag tataattatt 

tcgttatgaa atgggttaac
     7981 aaagaataca agaaaatatt tacaaaaaat caatttaaca 

attccttaaa acatgcagga
     8041 attgacgatt taaacaatat tagctttgaa caattcttat 

ctcttttcaa tagctataaa
     8101 ttatttaata agtaagttaa gggatgcata aactgcatcc 

cttaacttgt ttttcgtgta
     8161 cctatttttt gtgaatcgat tatgtctttt gcgcattcac 

ttcttttcta tataaatatg
     8221 agcgaagcga ataagcgtcg gaaaagcagc aaaaagtttc 

ctttttgctg ttggagcatg
     8281 ggggttcagg gggtgcagta tctgacgtca atgccgagcg 

aaagcgagcc gaagggtagc
     8341 atttacgtta gataaccccc tgatatgctc cgacgcttta 

tatagaaaag aagattcaac
     8401 taggtaaaat cttaatatag gttgagatga taaggtttat 

aaggaatttg tttgttctaa
     8461 tttttcactc attttgttct aatttctttt aacaaatgtt 

cttttttttt tagaacagtt
     8521 atgatatagt tagaatagtt taaaataagg agtgagaaaa 

agatgaaaga aagatatgga
     8581 acagtctata aaggctctca gaggctcata gacgaagaaa 

gtggagaagt catagaggta
     8641 gacaagttat accgtaaaca aacgtctggt aacttcgtaa 

aggcatatat agtgcaatta
     8701 ataagtatgt tagatatgat tggcggaaaa aaacttaaaa 

tcgttaacta tatcctagat
     8761 aatgtccact taagtaacaa tacaatgata gctacaacaa 

gagaaatagc aaaagctaca
     8821 ggaacaagtc tacaaacagt aataacaaca cttaaaatct 

tagaagaagg aaatattata
     8881 aaaagaaaaa ctggagtatt aatgttaaac cctgaactac 

taatgagagg cgacgaccaa
     8941 aaacaaaaat acctcttact cgaatttggg aactttgagc 

aagaggcaaa tgaaatagat
     9001 tgacctccca ataacaccac gtagttattg ggaggtcaat 

ctatgaaatg cgatta
//