LOCUS       sequence-388979-       11767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (Empty Backbone) Constitutive CRISPRi vector conferring hygromycin B
            resistance, ready for insertion of gRNA targeting gene of interest 
            using SapI restriction sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11767)
  AUTHORS   Czarnek M, Kochan J, Wawro M, Myrczek R, Bereta J
  TITLE     Construction of a Set of Novel Transposon Vectors for Efficient 
            Silencing of Protein and lncRNA Genes via CRISPR Interference.
  JOURNAL   Mol Biotechnol. 2023 Jan 28. doi: 10.1007/s12033-023-00675-5.
  PUBMED    36707469
REFERENCE   2  (bases 1 to 11767)
  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: "Mol 
            Biotechnol. 2023 Jan 28. doi: 10.1007/s12033-023-00675-5."
COMMENT     Sequence Label: pSBbi-Hyg-dCas9-KRAB-MeCP2_hU6-SapI
FEATURES             Location/Qualifiers
     source          1..11767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..1195
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     1143..1163
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1224..1233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1230..1256,1257..1259,1260..3743,3744..3749,3750..3815,
                     3816..3818,3819..5333)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5346..5366
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5427..5621
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             5655..5675
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5685..6542
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     primer_bind     complement(6596..6613)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6602..6826
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        7258..7362
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7363..7431,7432..8223)
                     /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(7581..7600)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8394..8982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8883..8902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9136..9153
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     9527..9546
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    9545..9600
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     9646..9665
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(9765..10790)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(9765..10790)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        11424..11672
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     11424..11444
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     11595..11614
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        11691..11766
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tttttgtttt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
       61 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
      121 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
      181 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
      241 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
      301 gtgccttgaa ttacttccac gcccctggct gcagtacgtg attcttgatc ccgagcttcg
      361 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
      421 ttgagttgag gcctggcttg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
      481 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
      541 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
      601 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
      661 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
      721 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
      781 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
      841 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
      901 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
      961 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     1021 ggggttttat gcgatggagt ttccccatac tgagtgggtg gagactgaag ttaggccagc
     1081 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     1141 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttaaggtgt cgtgaaaact
     1201 accccaagct ggcctctgag gccgccacca tggacaagaa gtactccatt gggctcgcta
     1261 tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa
     1321 aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc
     1381 tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca
     1441 gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg
     1501 ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata
     1561 aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa
     1621 agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact
     1681 tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg
     1741 agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga
     1801 cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa
     1861 tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg
     1921 gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca
     1981 actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct
     2041 acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt
     2101 tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg
     2161 agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag
     2221 acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt
     2281 tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg
     2341 aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg
     2401 taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc
     2461 cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc
     2521 cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact
     2581 atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag
     2641 agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct
     2701 tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac
     2761 actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca
     2821 cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc
     2881 tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa
     2941 agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc
     3001 tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg
     3061 agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga
     3121 tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc
     3181 tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc
     3241 gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc
     3301 ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag
     3361 cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc
     3421 cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa
     3481 tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc
     3541 agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac
     3601 tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc
     3661 tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca
     3721 atcggctctc cgactacgac gtggctgcta tcgtgcccca gtcttttctc aaagatgatt
     3781 ctattgataa taaagtgttg acaagatccg ataaagctag agggaagagt gataacgtcc
     3841 cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac
     3901 tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt
     3961 tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg
     4021 tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc
     4081 gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc
     4141 agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg
     4201 cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg
     4261 gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca
     4321 aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta
     4381 cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag
     4441 aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc
     4501 aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc
     4561 tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat
     4621 acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga
     4681 aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc
     4741 gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca
     4801 aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga
     4861 aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta
     4921 aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag
     4981 ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg
     5041 agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc
     5101 tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc
     5161 acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca
     5221 tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt
     5281 caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg
     5341 ctgaccccaa gaagaagagg aaggtgagtg gtggaggaag tggcgggtca gggtcgatgg
     5401 acgcgaaatc acttacggca tggtcgagaa cactggttac gttcaaggac gtgtttgtgg
     5461 actttacacg tgaggagtgg aaattgctgg atactgcgca acaaattgtg tatcgaaatg
     5521 tcatgcttga gaattacaag aacctcgtca gtctcggata ccagttgacg aaaccggatg
     5581 tgatccttag gctcgaaaag ggggaagaac cttggctggt atcgggaggt ggttcgggtg
     5641 gctctggatc aagcccaaag aagaaacgga aggtggaagc ctcagtgcag gtgaaaaggg
     5701 tgctggaaaa atcccccggc aaactcctcg tgaagatgcc cttccaggct tcccctggcg
     5761 gaaaaggtga agggggtggc gcaaccacat ctgcccaggt catggtcatc aagcgacctg
     5821 gaaggaaaag aaaggccgag gctgaccctc aggccattcc aaagaaacgg ggacgcaagc
     5881 cagggtccgt ggtcgcagct gcagcagctg aggctaagaa aaaggcagtg aaggaaagct
     5941 ccatccgcag tgtgcaggag actgtcctgc ccatcaagaa gaggaagact agggagaccg
     6001 tgtccatcga ggtcaaagaa gtggtcaagc ccctgctcgt gtccaccctg ggcgaaaaat
     6061 ctggaaaggg gctcaaaaca tgcaagtcac ctggacggaa aagcaaggag tctagtccaa
     6121 aggggcgctc aagctccgct tctagtcccc ctaaaaagga acaccatcac catcaccatc
     6181 acgccgagtc tcctaaggct cctatgccac tgctcccacc acctccacca cctgagccac
     6241 agtcaagcga agaccccatc agcccacccg agcctcagga tctgtcctct agtatttgca
     6301 aagaggaaaa gatgcccaga gcaggcagcc tggagagtga tggctgtcca aaagaacccg
     6361 ccaagaccca gcctatggtg gcagccgctg caactaccac cacaaccaca actaccacag
     6421 tggccgaaaa atacaagcat cgcggcgagg gcgaacgaaa ggacattgtg tcaagctcca
     6481 tgcccagacc taaccgggag gaaccagtcg atagtaggac acccgtgact gagagagtct
     6541 catagggcct gtcaggccaa gcttggggat caattctcta gagctcgctg atcagcctcg
     6601 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6661 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6721 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6781 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     6841 agaaccagct gggcctacta gatagcggac cccttaccga aacatcgccg cattctgcag
     6901 aggagtcgaa cttagtgtat gtaaacttct gacccactgg aattgtgata cagtgaatta
     6961 taagtgaaat aatctgtctg taaacaattg ttggaaaaat gacttgtgtc atgcacaaag
     7021 tagatgtcct aactgacttg ccaaaactat tgtttgttaa caagaaattt gtggagtagt
     7081 tgaaaaacga gttttaatga ctccaactta agtgtatgta aacttccgac ttcaactgta
     7141 tagggatcca attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     7201 ttacgcatct gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc
     7261 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     7321 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     7381 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     7441 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     7501 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7561 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     7621 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7681 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7741 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7801 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7861 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7921 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7981 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8041 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     8101 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     8161 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     8221 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     8281 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8341 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     8401 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     8461 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     8521 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     8581 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8641 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8701 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8761 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8821 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8881 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8941 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     9001 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     9061 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     9121 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg atccctatac agttgaagtc
     9181 ggaagtttac atacacctta gccaaataca tttaaactca ctttttcaca attcctgaca
     9241 tttaatccta gtaaaaattc cctgtcttag gtcagttagg atcaccactt tattttaaga
     9301 atgtgaaata tcagaataat agtagagaga atgattcatt tcagctttta tttctttcat
     9361 cacattccca gtgggtcaga agtttacata cactcgactc ctctgcagaa tgcggcgatg
     9421 tttcggtaag gggtcccgag tagctagttc atggcagcca gcatatggca tatgttgcca
     9481 aactctaaac caaatactca ttctgatgtt ttaaatgatt tgccctccca tatgtccttc
     9541 cgagtgagag acacaaaaaa ttccaacaca ctattgcaat gaaaataaat ttcctttatt
     9601 agccagaagt cagatgctca aggggcttca tgatgtcccc ataatttttg gcagagggaa
     9661 aaagatctca gtggtatttg tgagccaggg cattggccac accagccacc accttctgat
     9721 aggcagcctg cacctgagga gtgaattcac gacaggcctt cgaactattc ctttgccctc
     9781 ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc
     9841 cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg
     9901 acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag
     9961 ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct
    10021 gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac
    10081 ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc
    10141 cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc
    10201 gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga
    10261 gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg
    10321 ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt
    10381 ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc
    10441 tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca
    10501 ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc
    10561 acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag
    10621 aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg
    10681 aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt
    10741 tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttcat ggtggcctca
    10801 ggtgcagagg tttctacagg gaaagagaga aaggctacag ttagtttgct gtgatgatgc
    10861 taagtgccga ggtcagatgt ctatggattg tacacggaat cccaacgcag gaaccaaaaa
    10921 acactcaccc aaaaaaatgg cgctaaggcc gagagaaagg cgaagttccg tctacggcta
    10981 tataccttga aacaggggcg gcagaagtga cgcaacgcgg ttgcgcggac ggcttcctgt
    11041 gttttcattg gtcgaagcgt tctgcgcgtg cgcagagatt ccccagggaa ggcgggccct
    11101 cttccgaatg tccaatccgg acattcttcc ggttggacct ccgcggtcag cgctgtgccc
    11161 ggcgagagat cacgtggggc gcggaggcgg tgctgctggg gcacggccgt ccagcctcgg
    11221 cggccatatt tttgaggggc tgttcatctc gttcacacgc tctgtccgcc atgtttgtga
    11281 gtggaagcgc cattacccct tcaagcgact gaaggctgca gggcctctgg tggcccgcat
    11341 ggggagacca gacccgccag gcccgccttt ccgcactcag tccgggctta ctttattttg
    11401 tgagacaggg tctcgcctag agggagggcc tatttcccat gattccttca tatttgcata
    11461 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
    11521 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
    11581 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
    11641 ctttatatat cttgtggaaa ggacgaaaca ccggaagagc gagctcttct gttttagagc
    11701 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
    11761 cggtgct
//
