LOCUS       sequence-314821-       11839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobile CRISPRi sfGFP 'test', gmc6 sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11839)
  AUTHORS   Banta AB, Ward RD, Tran JS, Bacon EE, Peters JM
  TITLE     Programmable Gene Knockdown in Diverse Bacteria Using 
            Mobile-CRISPRi.
  JOURNAL   Curr Protoc Microbiol. 2020 Dec;59(1):e130. doi: 10.1002/cpmc.130.
  PUBMED    33332762
REFERENCE   2  (bases 1 to 11839)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr Protoc
            Microbiol."; date: "2020-12"; volume: "59(1):e130. doi"; pages: " 
            10.1002/cpmc.130"
COMMENT     Sequence Label: pJMP2774
FEATURES             Location/Qualifiers
     source          1..11839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             79..1161
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1446..1462
                     /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)."
     CDS             join(1818..1820,1821..2534)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1818..2531
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1848..1876)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2471..2491
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2676..2723)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        join(2840..2845,2846..2862,2863..2868)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3057..3590
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(3638..3685)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     terminator      complement(3736..3822)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3925..4019)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4047..4063)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4048..4064)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4478..4497)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4597..4619
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4657..4675)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4743..4847
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4848..4916,4917..5708)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5066..5085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6009..6397
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6666..6774
                     /note="incP origin of transfer"
     mobile_element  6878..7043
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7198..7287)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7297..8880,8881..8883,8884..11370,
                     11371..11373,11374..11400))
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     protein_bind    11442..11458
                     /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)."
ORIGIN
        1 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gatggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag atagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 accccacccg ggtccttgag ccccattatc acctccaaag ccttctcgtc tggtcagttt
     1261 cacctgtttt acgtaaaaac ccgcttcggc gggtttttac ttttggggaa acacagaaaa
     1321 aagcccgcac ctgacagtgc gggctttttt tttcgaccaa aggtgcgact actcttgcct
     1381 actacctatc gactgagctg aaagaattcc taaagatcta taaatgtgag cggataacat
     1441 tgacattgtg agcggataac aagatacttc tagtcatcta attcaacaag aattgtttaa
     1501 gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa
     1561 aagtggcacc gagtcggtgc tttttttttg aattcatgtg gctgaccgtt ctgttgtctc
     1621 tcgctcttcc gagtagacga acaataaggc ctccctaacg gggggccttt tttattgata
     1681 acaaaagtca gtgcttccgc tatttccaaa ataccgggct aatacggttt aaacgaaaat
     1741 ttatcaaaaa gagtattgac ttaaagtcta acctatagga tacttacagc cagatctgag
     1801 cgagaaggag gtaaagtatg agcaaaggag aagaactttt cactggagtt gtcccaattc
     1861 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtccgtgga gagggtgaag
     1921 gtgatgctac aaacggaaaa ctcaccctta aatttatttg cactactgga aaactacctg
     1981 ttccgtggcc aacacttgtc actactctga cctatggtgt tcaatgcttt tcccgttatc
     2041 cggatcacat gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg
     2101 aacgcactat atctttcaaa gatgacggga cctacaagac gcgtgctgaa gtcaagtttg
     2161 aaggtgatac ccttgttaat cgtatcgagt taaagggtat tgattttaaa gaagatggaa
     2221 acattcttgg acacaaactg gagtacaact ttaactcaca caatgtatac atcacggcag
     2281 acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacgtt gaagatggtt
     2341 ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     2401 taccagacaa ccattacctg tcgacacaat ctgtcctttc gaaagatccc aacgaaaagc
     2461 gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg
     2521 agctctacaa ataataagtt taaacgacct cctggatttg ctcagacagc cttttcgtca
     2581 ttcgtttcag ccaaaaaact taagaccgcc ggtcttgtcc actaccttgc agtaatgcgg
     2641 tggacaggat cggcggtttt cttttctctt ctcaagaagt tcctatactt tctagagaat
     2701 aggaacttcg gaataggaac ttcctcctga acggccataa gaacgaaggc tgtctgttga
     2761 actctcgagc cttgtcgcct tgcgtataat atttgcccat ggacgcacac cgtggaaacg
     2821 gatgaaggca cgaacccagt tgacataagc ctgttcggtt cgtaaactgt aatgcaagta
     2881 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     2941 agtggcggtt ttcatggctt gttatgactg tttttttgta cagtctatgc ctcgggcatc
     3001 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     3061 tacgcagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaggtggct
     3121 caagtatggg catcattcgc acatgtaggc tcggccctga ccaagtcaaa tccatgcggg
     3181 ctgctcttga tcttttcggt cgtgagttcg gagacgtagc cacctactcc caacatcagc
     3241 cggactccga ttacctcggg aacttgctcc gtagtaagac attcatcgcg cttgctgcct
     3301 tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt tctgcccaag tttgagcagc
     3361 cgcgtagtga gatctatatc tatgatctcg cagtctccgg agagcaccgg aggcagggca
     3421 ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga
     3481 tctacgtgca agcagattac ggtgacgatc ccgcagtggc tctctataca aagttgggca
     3541 tacgggaaga agtgatgcac tttgatatcg acccaagtac cgccacctaa caattcgttc
     3601 aagccgagat ctcgagcatc agcttcaaaa gcgctctgaa gttcctatac tttctagaga
     3661 ataggaactt cggaataggt acttcaagat ccccaattcg agctcatcgt ccgggccgca
     3721 agctcctagc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     3781 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctca agctagagag
     3841 tcattacccc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     3901 gccactcatc gcagtctagc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     3961 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     4021 aagctcagct aattaagcta gcttatcgat accgtcgacc tcgaacccca cgcccctctt
     4081 taatacgacg ggcaatttgc acttcagaaa atgaagagtt tgctttagcc ataacaaaag
     4141 tccagtatgc tttttcacag cataactgga ctgatttcag tttacaacta ttctgtctag
     4201 tttaagactt tattgtcata gtttagatct attttgttca gtttaagact ttattgtccg
     4261 cccacacctg cttacgcagg gcatccattt attactcaac cgtaaccgat tttgccaggt
     4321 tacgcggctg gtcctctagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4381 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     4441 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     4501 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4561 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4621 tttcaccgtc gtcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4681 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4741 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4801 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4861 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4921 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4981 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5041 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5101 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5161 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5221 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5281 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5341 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5401 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5461 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5521 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcccgc ggtatcattg
     5581 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5641 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5701 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5761 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5821 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggcagttatt
     5881 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     5941 attgatttaa aacttcattt ttaatttttg cggccgcaag atccggccac gatgcgtccg
     6001 gcgtagagga tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg
     6061 tttcacgtac taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc
     6121 ctcatggcta acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac
     6181 taagctctca tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag
     6241 gttttaagtt ttataagaaa aaaaagaata tataaggctt ttaaagcttt taaggtttaa
     6301 cggttgtgga caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag
     6361 caattttgag tgacacagga acacttaacg gctgacatgg gaattagcca tggcatcaca
     6421 gtatcgtgat gacagaggca gggagtgcgg cctttttacg gttcctggcc ttttgctggc
     6481 cttttgctca catgttcttt cctgcgttat caggggattc cttaaggtat actttccgct
     6541 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
     6601 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
     6661 cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc
     6721 ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccgataagc
     6781 tctgataccg ctcgccgcag ccgaacgacc gagcgcagcg ggtcagtgag cgaggaagcg
     6841 gaagagccca aacataacag gaagaaaaat gccccgctgt gggcggacaa aatagttggg
     6901 aactgggagg ggtggaaatg gagtttttaa ggattattta gggaagagtg acaaaataga
     6961 tgggaactgg gtgtagcgtc gtaagctaat acgaaaatta aaaatgacaa aatagtttgg
     7021 aactagattt cacttatctg gttcttgagg cggttaaaag agccgtactc ttctccgatg
     7081 tcgactaggc catgatgctc attctgtggg accaaaacga aaaaacaccc tttcgggtgt
     7141 cttttctgga atttggtacc gaggctgcaa gtcccattaa ggaggcgcgc cttattacag
     7201 atcttcctca gagatgagct tctgttccag atcttcttct gaaatcaact tttgttccag
     7261 atcttcttca gagatgagtt tctgctccgc ggccgcgtca cctcctagct gactcaaatc
     7321 aatgcgtgtt tcataaagac cagtgatgga ttgatggata agagtggcat ctaaaacttc
     7381 ttttgtagac gtatatcgtt tacgatcaat tgttgtatca aaatatttaa aagcagcggg
     7441 agctccaaga ttcgtcaacg taaataaatg aataatattt tctgcttgtt cacgtattgg
     7501 tttgtctcta tgtttgttat atgcactaag aactttatct aaattggcat ctgctaaaat
     7561 aacacgctta gaaaattcac tgatttgctc aataatctca tctaaataat gcttatgctg
     7621 ctccacaaac aattgttttt gttcgttatc ttctggacta cccttcaact tttcataatg
     7681 actagctaaa tataaaaaat tcacatattt gcttggcaga gccagctcat ttcctttttg
     7741 taattctccg gcactagcca gcatccgttt acgaccgttt tctaactcaa aaagactata
     7801 tttaggtagt ttaatgatta agtctttttt aacttcctta tatcctttag cttctaaaaa
     7861 gtcaatcgga tttttttcaa aggaacttct ttccataatt gtgatcccta gtaactcttt
     7921 aacggatttt aacttcttcg atttcccttt ttccacctta gcaaccacta ggactgaata
     7981 agctaccgtt ggactatcaa aaccaccata tttttttgga tcccagtctt ttttacgagc
     8041 aataagcttg tccgaatttc tttttggtaa aattgactcc ttggagaatc cgcctgtctg
     8101 tacttctgtt ttcttgacaa tattgacttg gggcatggac aatactttgc gcactgtggc
     8161 aaaatctcgc cctttatccc agacaatttc tccagtttcc ccattagttt cgattagagg
     8221 gcgtttgcga atctctccat ttgcaagtgt aatttctgtt ttgaagaagt tcatgatatt
     8281 agagtaaaag aaatattttg cggttgcttt gcctatttct tgctcagact tagcaatcat
     8341 tttacgaaca tcataaactt tataatcacc atagacaaac tccgattcaa gttttggata
     8401 tttcttaatc aaagcagttc caacgacggc atttagatac gcatcatggg catgatggta
     8461 attgttaatc tcacgtactt tatagaattg gaaatctttt cggaagtcag aaactaattt
     8521 agattttaag gtaatcactt taacctctcg aataagttta tcattttcat cgtatttagt
     8581 attcatgcga ctatccaaaa tttgtgccac atgcttagtg atttggcgag tttcaaccaa
     8641 ttggcgtttg ataaaaccag ctttatcaag ttcactcaaa cctccacgtt cagctttcgt
     8701 taaattatca aacttacgtt gagtgattaa cttggcgttt agaagttgtc tccaatagtt
     8761 tttcatcttt ttgactactt cttcacttgg aacgttatcc gatttaccac gatttttatc
     8821 agaacgcgtt aagaccttat tgtctattga atcgtcttta aggaaacttt gtggaacaat
     8881 ggcatcgaca tcataatcac ttaaacgatt aatatctaat tcttggtcca catacatgtc
     8941 tcttccattt tggagataat agagatagag cttttcattt tgcaattgag tattttcaac
     9001 aggatgctct ttaagaatct gacttcctaa ttctttgata ccttcttcga ttcgtttcat
     9061 acgctctcgc gaatttttct ggcccttttg agttgtctga ttttcacgtg ccatttcaat
     9121 aacgatattt tctggcttat gccgccccat tactttgacc aattcatcaa caacttttac
     9181 agtctgtaaa ataccttttt taatagcagg gctaccagct aaatttgcaa tatgttcatg
     9241 taaactatcg ccttgtccag acacttgtgc tttttgaata tcttctttaa atgtcaaact
     9301 atcatcatgg atcagctgca taaaattgcg attggcaaaa ccatctgatt tcaaaaaatc
     9361 taatattgtt ttgccagatt gcttatccct aataccatta atcaattttc gagacaaacg
     9421 tccccaacca gtataacggc gacgtttaag ctgtttcatc accttatcat caaagaggtg
     9481 agcatatgtt ttaagtcttt cctcaatcat ctccctatct tcaaataagg tcaatgttaa
     9541 aacaatatcc tctaagatat cttcattttc ttcattatcc aaaaaatctt tatctttaat
     9601 aatttttagc aaatcatggt aggtacctaa tgaagcatta aatctatctt caactcctga
     9661 aatttcaaca ctatcaaaac attctatttt tttgaaataa tcttctttta attgcttaac
     9721 ggttactttt cgatttgttt tgaagagtaa atcaacaatg gctttcttct gttcacctga
     9781 aagaaatgct ggttttcgca ttccttcagt aacatatttg acctttgtca attcgttata
     9841 aaccgtaaaa tactcataaa gcaaactatg ttttggtagt actttttcat ttggaagatt
     9901 tttatcaaag tttgtcatgc gttcaataaa tgattgagct gaagcacctt tatcgacaac
     9961 ttcttcaaaa ttccatgggg taattgtttc ttcagacttc cgagtcatcc atgcaaaacg
    10021 actattgcca cgcgccaatg gaccaacata ataagggatt cgaaaagtca agattttttc
    10081 aatcttctca cgattgtctt ttaaaaatgg ataaaaatct tcttgtcgtc tcaaaatagc
    10141 atgcagctca cccaagtgaa tttgatgggg aatagagccg ttgtcaaagg tccgttgctt
    10201 gcgcagcaaa tcttcacgat ttagtttcac caataattcc tcagtaccat ccattttttc
    10261 taaaattggt ttgataaatt tataaaattc ttcttggcta gctcccccat caatataacc
    10321 tgcatatccg ttttttgatt gatcaaaaaa gatttcttta tacttttctg gaagttgttg
    10381 tcgaactaaa gcttttaaaa gagtcaagtc ttgatgatgt tcatcgtagc gtttaatcat
    10441 tgaagctgat aggggagcct tagttatttc agtatttact cttaggatat ctgaaagtaa
    10501 aatagcatct gataaattct tagctgccaa aaacaaatca gcatattgat ctccaatttg
    10561 cgccaataaa ttatctaaat catcatcgta agtatctttt gaaagctgta atttagcatc
    10621 ttctgccaaa tcaaaatttg atttaaaatt aggggtcaaa cccaatgaca aagcaatgag
    10681 attcccaaat aagccatttt tcttctcacc ggggagctga gcaatgagat tttctaatcg
    10741 tcttgattta ctcaatcgtg cagaaagaat cgctttagca tctactccac ttgcgttaat
    10801 agggttttct tcaaataatt gattgtaggt ttgtaccaac tggataaata gtttgtccac
    10861 atcactatta tcaggattta aatctccctc aatcaaaaaa tgaccacgaa acttaatcat
    10921 atgcgctaag gccaaataga ttaagcgcaa atccgcttta tcagtagaat ctaccaattt
    10981 ttttcgcaga tgatagatag ttggatattt ctcatgataa gcaacttcat ctactatatt
    11041 tccaaaaata ggatgacgtt catgcttctt atcttcttcc accaaaaaag actcttcaag
    11101 tcgatgaaag aaactatcat ctactttcgc catctcattt gaaaaaatct cctgtagata
    11161 acaaatacga ttcttccgac gtgtatacct tctacgagct gtccgtttga gacgagtcgc
    11221 ttccgctgtc tctccactgt caaataaaag agcccctata agattttttt tgatactgtg
    11281 gcggtctgta tttcccagaa ccttgaactt tttagacgga accttatatt catcagtgat
    11341 caccgcccat ccgacgctat ttgtgccgat agctaagcct attgagtatt tcttatccat
    11401 actagttttc ctgtgtgaaa cctgctgatg tgctcagtat cttgttatcc gctcacaatg
    11461 tcaatgttat ccgctcacat ttataatatt ttatctgatt aataagatga tcttcttgag
    11521 atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg
    11581 gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggtgata
    11641 agctgtcaaa ccagatcaat tcgcactagt ctgcagacgt aaaaaaagcg gcgtggttag
    11701 ccgctttttt aattgccgga gcgcaataaa aaagcccccg gaaggtgatc ttccgggggc
    11761 tttctcatgc gttgcttgta ttgaaaatgg gggccatgcc tacctatctg cctcccggga
    11821 agcggcatgc atttacgtt
//
