LOCUS       sequence-474448-       15788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Tet1CD-dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15788)
  AUTHORS   Wu Y, Zhong A, Evangelisti A, Sidharta M, Danwei H, Studer L, Zhou T
  TITLE     Leveraging CRISPR activation for rapid assessment of gene editing 
            products in human pluripotent stem cells.
  JOURNAL   Stem Cell Reports. 2025 Apr 25:102499. doi: 
            10.1016/j.stemcr.2025.102499.
  PUBMED    40345204
REFERENCE   2  (bases 1 to 15788)
  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: "Stem Cell 
            Reports. 2025 Apr 25:102499. doi: 10.1016/j.stemcr.2025.102499."
COMMENT     Sequence Label: pLV-TET1-dCas9
FEATURES             Location/Qualifiers
     source          1..15788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..30
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(149..169)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        154..483
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      334..469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     396..415
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        531..578
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             597..971
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1101..1222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1138..1157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1192..1211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1271..1287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1271..1287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1284..1306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1295..1311
                     /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)."
     promoter        complement(join(1319..1325,1326..1343,1344..1349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1364..1385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1502..1519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1673..2261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1753..1772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2432..3223,3224..3292))
                     /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     3055..3074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3293..3397)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3472..3491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3667..4046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4048..4246
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4201..4221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4264..4444
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4491..4616
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5109..5342
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5527..5571
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5869..5986
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6037..7218
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6267..7209
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7166..7186
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7228..7237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(9634..9660,9661..9663,9664..12150,12151..12153,
                     12154..13737)
                     /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"
     CDS             13753..13773
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13777..13797
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13801..13821
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13834..13890
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             13891..14289
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    14305..14893
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(14358..14378)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(14776..14787)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             14965..15198
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(15224..15241)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    15230..15454
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(15500..15788,1..140)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(15587..15606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
       61 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      121 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
      241 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      301 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      361 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      421 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      481 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
      541 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
      601 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
      661 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
      721 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
      781 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
      841 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
      901 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
      961 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     1021 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     1081 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     1141 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1201 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     1261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1321 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1861 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2101 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3421 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     3481 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     3541 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     3601 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     3661 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     3721 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3781 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3841 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3901 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     3961 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4021 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4081 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4141 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4201 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
     4261 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     4321 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     4381 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     4441 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     4501 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     4561 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     4621 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     4681 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     4741 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     4801 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     4861 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     4921 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     4981 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     5041 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     5101 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     5161 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     5221 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     5281 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     5341 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     5401 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     5461 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     5521 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     5581 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     5641 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     5701 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     5761 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     5821 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     5881 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     5941 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     6001 ggacagcaga gatccagttt ggttaattaa ggtaccgctc cggtgcccgt cagtgggcag
     6061 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     6121 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     6181 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     6241 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     6301 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacgcccctg gctgcagtac
     6361 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     6421 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     6481 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     6541 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     6601 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     6661 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     6721 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     6781 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     6841 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     6901 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     6961 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     7021 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     7081 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     7141 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     7201 ccatttcagg tgtcgtgaga attctgcgcc accatggccc tgcccacctg cagctgtctt
     7261 gatcgagtta tacaaaaaga caaaggccca tattatacac accttggggc aggaccaagt
     7321 gttgctgctg tcagggaaat catggagaat aggtatggtc aaaaaggaaa cgcaataagg
     7381 atagaaatag tagtgtacac cggtaaagaa gggaaaagct ctcatgggtg tccaattgct
     7441 aagtgggttt taagaagaag cagtgatgaa gaaaaagttc tttgtttggt ccggcagcgt
     7501 acaggccacc actgtccaac tgctgtgatg gtggtgctca tcatggtgtg ggatggcatc
     7561 cctcttccaa tggccgaccg gctatacaca gagctcacag agaatctaaa gtcatacaat
     7621 gggcacccta ccgacagaag atgcaccctc aatgaaaatc gtacctgtac atgtcaagga
     7681 attgatccag agacttgtgg agcttcattc tcttttggct gttcatggag tatgtacttt
     7741 aatggctgta agtttggtag aagcccaagc cccagaagat ttagaattga tccaagctct
     7801 cccttacatg aaaaaaacct tgaagataac ttacagagtt tggctacacg attagctcca
     7861 atttataagc agtatgctcc agtagcttac caaaatcagg tggaatatga aaatgttgcc
     7921 cgagaatgtc ggcttggcag caaggaaggt cgacccttct ctggggtcac tgcttgcctg
     7981 gacttctgtg ctcatcccca cagggacatt cacaacatga ataatggaag cactgtggtt
     8041 tgtaccttaa ctcgagaaga taaccgctct ttgggtgtta ttcctcaaga tgagcagctc
     8101 catgtgctac ctctttataa gctttcagac acagatgagt ttggctccaa ggaaggaatg
     8161 gaagccaaga tcaaatctgg ggccatcgag gtcctggcac cccgccgcaa aaaaagaacg
     8221 tgtttcactc agcctgttcc ccgttctgga aagaagaggg ctgcgatgat gacagaggtt
     8281 cttgcacata agataagggc agtggaaaag aaacctattc cccgaatcaa gcggaagaat
     8341 aactcaacaa caacaaacaa cagtaagcct tcgtcactgc caaccttagg gagtaacact
     8401 gagaccgtgc aacctgaagt aaaaagtgaa accgaacccc attttatctt aaaaagttca
     8461 gacaacacta aaacttattc gctgatgcca tccgctcctc acccagtgaa agaggcatct
     8521 ccaggcttct cctggtcccc gaagactgct tcagccacac cagctccact gaagaatgac
     8581 gcaacagcct catgcgggtt ttcagaaaga agcagcactc cccactgtac gatgccttcg
     8641 ggaagactca gtggtgccaa tgctgcagct gctgatggcc ctggcatttc acagcttggc
     8701 gaagtggctc ctctccccac cctgtctgct cctgtgatgg agcccctcat taattctgag
     8761 ccttccactg gtgtgactga gccgctaacg cctcatcagc caaaccacca gccctccttc
     8821 ctcacctctc ctcaagacct tgcctcttct ccaatggaag aagatgagca gcattctgaa
     8881 gcagatgagc ctccatcaga cgaaccccta tctgatgacc ccctgtcacc tgctgaggag
     8941 aaattgcccc acattgatga gtattggtca gacagtgagc acatcttttt ggatgcaaat
     9001 attggtgggg tggccatcgc acctgctcac ggctcggttt tgattgagtg tgcccggcga
     9061 gagctgcacg ctaccactcc tgttgagcac cccaaccgta atcatccaac ccgcctctcc
     9121 cttgtctttt accagcacaa aaacctaaat aagccccaac atggttttga actaaacaag
     9181 attaagtttg aggctaaaga agctaagaat aagaaaatga aggcctcaga gcaaaaagac
     9241 caggcagcta atgaaggtcc agaacagtcc tctgaagtaa atgaattgaa ccaaattcct
     9301 tctcataaag cattaacatt aacccatgac aatgttgtca ccgtgtcccc ttatgctctc
     9361 acacacgttg cggggcccta taaccattgg gtcggagggc cgagctctgg cgcaccccca
     9421 ccaagtggag ggtctcctgc cgggtcccca acatctactg aagaaggcac cagcgaatcc
     9481 gcaacgcccg agtcaggccc tggtacctcc acagaaccat ctgaaggtag tgcgcctggt
     9541 tccccagctg gaagccctac ttccaccgaa gaaggcacgt caaccgaacc aagtgaagga
     9601 tctgcccctg ggaccagcac tgaaccatct gagatggaca agaagtattc tatcggactg
     9661 gccatcggga ctaatagcgt cgggtgggcc gtgatcactg acgagtacaa ggtgccctct
     9721 aagaagttca aggtgctcgg gaacaccgac cggcattcca tcaagaaaaa tctgatcgga
     9781 gctctcctct ttgattcagg ggagaccgct gaagcaaccc gcctcaagcg gactgctaga
     9841 cggcggtaca ccaggaggaa gaaccggatt tgttaccttc aagagatatt ctccaacgaa
     9901 atggcaaagg tcgacgacag cttcttccat aggctggaag aatcattcct cgtggaagag
     9961 gataagaagc atgaacggca tcccatcttc ggtaatatcg tcgacgaggt ggcctatcac
    10021 gagaaatacc caaccatcta ccatcttcgc aaaaagctgg tggactcaac cgacaaggca
    10081 gacctccggc ttatctacct ggccctggcc cacatgatca agttcagagg ccacttcctg
    10141 atcgagggcg acctcaatcc tgacaatagc gatgtggata aactgttcat ccagctggtg
    10201 cagacttaca accagctctt tgaagagaac cccatcaatg caagcggagt cgatgccaag
    10261 gccattctgt cagcccggct gtcaaagagc cgcagacttg agaatcttat cgctcagctg
    10321 ccgggtgaaa agaaaaatgg actgttcggg aacctgattg ctctttcact tgggctgact
    10381 cccaatttca agtctaattt cgacctggca gaggatgcca agctgcaact gtccaaggac
    10441 acctatgatg acgatctcga caacctcctg gcccagatcg gtgaccaata cgccgacctt
    10501 ttccttgctg ctaagaatct ttctgacgcc atcctgctgt ctgacattct ccgcgtgaac
    10561 actgaaatca ccaaggcccc tctttcagct tcaatgatta agcggtatga tgagcaccac
    10621 caggacctga ccctgcttaa ggcactcgtc cggcagcagc ttccggagaa gtacaaggaa
    10681 atcttctttg accagtcaaa gaatggatac gccggctaca tcgacggagg tgcctcccaa
    10741 gaggaatttt ataagtttat caaacctatc cttgagaaga tggacggcac cgaagagctc
    10801 ctcgtgaaac tgaatcggga ggatctgctg cggaagcagc gcactttcga caatgggagc
    10861 attccccacc agatccatct tggggagctt cacgccatcc ttcggcgcca agaggacttc
    10921 tacccctttc ttaaggacaa cagggagaag attgagaaaa ttctcacttt ccgcatcccc
    10981 tactacgtgg gacccctcgc cagaggaaat agccggtttg cttggatgac cagaaagtca
    11041 gaagaaacta tcactccctg gaacttcgaa gaggtggtgg acaagggagc cagcgctcag
    11101 tcattcatcg aacggatgac taacttcgat aagaacctcc ccaatgagaa ggtcctgccg
    11161 aaacattccc tgctctacga gtactttacc gtgtacaacg agctgaccaa ggtgaaatat
    11221 gtcaccgaag ggatgaggaa gcccgcattc ctgtcaggcg aacaaaagaa ggcaattgtg
    11281 gaccttctgt tcaagaccaa tagaaaggtg accgtgaagc agctgaagga ggactatttc
    11341 aagaaaattg aatgcttcga ctctgtggag attagcgggg tcgaagatcg gttcaacgca
    11401 agcctgggta cctaccatga tctgcttaag atcatcaagg acaaggattt tctggacaat
    11461 gaggagaacg aggacatcct tgaggacatt gtcctgactc tcactctgtt cgaggaccgg
    11521 gaaatgatcg aggagaggct taagacctac gcccatctgt tcgacgataa agtgatgaag
    11581 caacttaaac ggagaagata taccggatgg ggacgcctta gccgcaaact catcaacgga
    11641 atccgggaca aacagagcgg aaagaccatt cttgatttcc ttaaaagcga cggattcgct
    11701 aatcgcaact tcatgcaact tatccatgat gattccctga cctttaagga ggacatccag
    11761 aaggcccaag tgtctggaca aggtgactca ctgcacgagc atatcgcaaa tctggctggt
    11821 tcacccgcta ttaagaaggg tattctccag accgtgaaag tcgtggacga gctggtcaag
    11881 gtgatgggtc gccataaacc agagaacatt gtcatcgaga tggccaggga aaaccagact
    11941 acccagaagg gacagaagaa cagcagggag cggatgaaaa gaattgagga agggattaag
    12001 gagctcgggt cacagatcct taaagagcac ccggtggaaa acacccagct tcagaatgag
    12061 aagctctatc tgtactacct tcaaaatgga cgcgatatgt atgtggacca agagcttgat
    12121 atcaacaggc tctcagacta cgacgtggac gccatcgtcc ctcagagctt cctcaaagac
    12181 gactcaattg acaataaggt gctgactcgc tcagacaaga accggggaaa gtcagataac
    12241 gtgccctcag aggaagtcgt gaaaaagatg aagaactatt ggcgccagct tctgaacgca
    12301 aagctgatca ctcagcggaa gttcgacaat ctcactaagg ctgagagggg cggactgagc
    12361 gaactggaca aagcaggatt cattaaacgg caacttgtgg agactcggca gattactaaa
    12421 catgtcgccc aaatccttga ctcacgcatg aataccaagt acgacgaaaa cgacaaactt
    12481 atccgcgagg tgaaggtgat taccctgaag tccaagctgg tcagcgattt cagaaaggac
    12541 tttcaattct acaaagtgcg ggagatcaat aactatcatc atgctcatga cgcatatctg
    12601 aatgccgtgg tgggaaccgc cctgatcaag aagtacccaa agctggaaag cgagttcgtg
    12661 tacggagact acaaggtcta cgacgtgcgc aagatgattg ccaaatctga gcaggagatc
    12721 ggaaaggcca ccgcaaagta cttcttctac agcaacatca tgaatttctt caagaccgaa
    12781 atcacccttg caaacggtga gatccggaag aggccgctca tcgagactaa tggggagact
    12841 ggcgaaatcg tgtgggacaa gggcagagat ttcgctaccg tgcgcaaagt gctttctatg
    12901 cctcaagtga acatcgtgaa gaaaaccgag gtgcaaaccg gaggcttttc taaggaatca
    12961 atcctcccca agcgcaactc cgacaagctc attgcaagga agaaggattg ggaccctaag
    13021 aagtacggcg gattcgattc accaactgtg gcttattctg tcctggtcgt ggctaaggtg
    13081 gaaaaaggaa agtctaagaa gctcaagagc gtgaaggaac tgctgggtat caccattatg
    13141 gagcgcagct ccttcgagaa gaacccaatt gactttctcg aagccaaagg ttacaaggaa
    13201 gtcaagaagg accttatcat caagctccca aagtatagcc tgttcgaact ggagaatggg
    13261 cggaagcgga tgctcgcctc cgctggcgaa cttcagaagg gtaatgagct ggctctcccc
    13321 tccaagtacg tgaatttcct ctaccttgca agccattacg agaagctgaa ggggagcccc
    13381 gaggacaacg agcaaaagca actgtttgtg gagcagcata agcattatct ggacgagatc
    13441 attgagcaga tttccgagtt ttctaaacgc gtcattctcg ctgatgccaa cctcgataaa
    13501 gtccttagcg catacaataa gcacagagac aaaccaattc gggagcaggc tgagaatatc
    13561 atccacctgt tcaccctcac caatcttggt gcccctgccg cattcaagta cttcgacacc
    13621 accatcgacc ggaaacgcta tacctccacc aaagaagtgc tggacgccac cctcatccac
    13681 cagagcatca ccggacttta cgaaactcgg attgacctct cacagctcgg aggggatggt
    13741 ggcggaggct cgccaaaaaa gaagagaaag gtagacccaa agaaaaaacg aaaagtagat
    13801 ccgaaaaaga agaggaaggt gggaggtagt ggggctacta acttcagcct gctgaagcag
    13861 gctggagacg tggaggagaa ccctggacct atggccaagc ctttgtctca agaagaatcc
    13921 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
    13981 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
    14041 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
    14101 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
    14161 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
    14221 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
    14281 gagggctaaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
    14341 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
    14401 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
    14461 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
    14521 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
    14581 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
    14641 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
    14701 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
    14761 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
    14821 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
    14881 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
    14941 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
    15001 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
    15061 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
    15121 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
    15181 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
    15241 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
    15301 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
    15361 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
    15421 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
    15481 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
    15541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
    15601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
    15661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
    15721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
    15781 tccacgtt
//
