LOCUS       sequence-467826-       11642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet1CD fused to N-terminus of dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11642)
  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 11642)
  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: EF1a-TET1-dCas9
FEATURES             Location/Qualifiers
     source          1..11642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1907..2605)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             complement(2606..2662)
                     /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             complement(2675..2695)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2699..2719)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2723..2743)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(2759..4342,4343..4345,4346..6832,6833..6835,
                     6836..6862))
                     /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"
     regulatory      9259..9268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9278..10456)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          9287..10225
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(9310..9330)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        10507..10611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10612..10680,10681..11472)
                     /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(10830..10849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagcca
     1861 ccaccttctg ataggcagcc tgcacctgag gagtgcggcc gctttactaa ttaagcttgt
     1921 gccccagttt gctagggagg tcgcagtatc tggccactgc cacctcgtgc tgctcgacgt
     1981 aggtctcgtt gttggcctcc ttgattcttt ccagtctgta gtccacatag tagacgccag
     2041 gcatcttgag gttcttagcg ggtttcttgg atctatatgt ggtcttgatg tttgcgatca
     2101 gatggctccc gcccacgagc ttcagggcca tgtcgtttct gccttccagg ccgccgtcag
     2161 cggggtacag cgtctcggtg aaggcctccc agccgagtgt tttcttctgc atcacagggc
     2221 cgttggatgt gaagttcacc cctctgatct tgacgttgta gatgaggcag ccgtcctgga
     2281 ggctggtgtc ctgggtagcg gtcagcacgc ccccgtcttc gtatgtggtg actctctccc
     2341 atgtgaagcc ctcagggaag gactgcttga agaagtcggg gatgccctgg gtgtggttga
     2401 tgaaggtctt gctgccgtag aggaagctag tagccaggat gtcgaaggcg aaggggagag
     2461 ggccgccctc gaccaccttg attctcatgg tctgggtgcc ctcgtagggc ttgccttcgc
     2521 cctcggatgt gcacttgaag tgatggttgt ccacggtgcc ctccatgtac agcttcatgt
     2581 gcatgttctc cttaatcagc tcgctaggtc cagggttctc ctccacgtct ccagcctgct
     2641 tcagcaggct gaagttagta gccccactac ctcccacctt cctcttcttt ttcggatcta
     2701 cttttcgttt tttctttggg tctacctttc tcttcttttt tggcgagcct ccgccaccat
     2761 cccctccgag ctgtgagagg tcaatccgag tttcgtaaag tccggtgatg ctctggtgga
     2821 tgagggtggc gtccagcact tctttggtgg aggtatagcg tttccggtcg atggtggtgt
     2881 cgaagtactt gaatgcggca ggggcaccaa gattggtgag ggtgaacagg tggatgatat
     2941 tctcagcctg ctcccgaatt ggtttgtctc tgtgcttatt gtatgcgcta aggactttat
     3001 cgaggttggc atcagcgaga atgacgcgtt tagaaaactc ggaaatctgc tcaatgatct
     3061 cgtccagata atgcttatgc tgctccacaa acagttgctt ttgctcgttg tcctcggggc
     3121 tccccttcag cttctcgtaa tggcttgcaa ggtagaggaa attcacgtac ttggagggga
     3181 gagccagctc attacccttc tgaagttcgc cagcggaggc gagcatccgc ttccgcccat
     3241 tctccagttc gaacaggcta tactttggga gcttgatgat aaggtccttc ttgacttcct
     3301 tgtaaccttt ggcttcgaga aagtcaattg ggttcttctc gaaggagctg cgctccataa
     3361 tggtgatacc cagcagttcc ttcacgctct tgagcttctt agactttcct ttttccacct
     3421 tagccacgac caggacagaa taagccacag ttggtgaatc gaatccgccg tacttcttag
     3481 ggtcccaatc cttcttcctt gcaatgagct tgtcggagtt gcgcttgggg aggattgatt
     3541 ccttagaaaa gcctccggtt tgcacctcgg ttttcttcac gatgttcact tgaggcatag
     3601 aaagcacttt gcgcacggta gcgaaatctc tgcccttgtc ccacacgatt tcgccagtct
     3661 ccccattagt ctcgatgagc ggcctcttcc ggatctcacc gtttgcaagg gtgatttcgg
     3721 tcttgaagaa attcatgatg ttgctgtaga agaagtactt tgcggtggcc tttccgatct
     3781 cctgctcaga tttggcaatc atcttgcgca cgtcgtagac cttgtagtct ccgtacacga
     3841 actcgctttc cagctttggg tacttcttga tcagggcggt tcccaccacg gcattcagat
     3901 atgcgtcatg agcatgatga tagttattga tctcccgcac tttgtagaat tgaaagtcct
     3961 ttctgaaatc gctgaccagc ttggacttca gggtaatcac cttcacctcg cggataagtt
     4021 tgtcgttttc gtcgtacttg gtattcatgc gtgagtcaag gatttgggcg acatgtttag
     4081 taatctgccg agtctccaca agttgccgtt taatgaatcc tgctttgtcc agttcgctca
     4141 gtccgcccct ctcagcctta gtgagattgt cgaacttccg ctgagtgatc agctttgcgt
     4201 tcagaagctg gcgccaatag ttcttcatct ttttcacgac ttcctctgag ggcacgttat
     4261 ctgactttcc ccggttcttg tctgagcgag tcagcacctt attgtcaatt gagtcgtctt
     4321 tgaggaagct ctgagggacg atggcgtcca cgtcgtagtc tgagagcctg ttgatatcaa
     4381 gctcttggtc cacatacata tcgcgtccat tttgaaggta gtacagatag agcttctcat
     4441 tctgaagctg ggtgttttcc accgggtgct ctttaaggat ctgtgacccg agctccttaa
     4501 tcccttcctc aattcttttc atccgctccc tgctgttctt ctgtcccttc tgggtagtct
     4561 ggttttccct ggccatctcg atgacaatgt tctctggttt atggcgaccc atcaccttga
     4621 ccagctcgtc cacgactttc acggtctgga gaataccctt cttaatagcg ggtgaaccag
     4681 ccagatttgc gatatgctcg tgcagtgagt caccttgtcc agacacttgg gccttctgga
     4741 tgtcctcctt aaaggtcagg gaatcatcat ggataagttg catgaagttg cgattagcga
     4801 atccgtcgct tttaaggaaa tcaagaatgg tctttccgct ctgtttgtcc cggattccgt
     4861 tgatgagttt gcggctaagg cgtccccatc cggtatatct tctccgttta agttgcttca
     4921 tcactttatc gtcgaacaga tgggcgtagg tcttaagcct ctcctcgatc atttcccggt
     4981 cctcgaacag agtgagagtc aggacaatgt cctcaaggat gtcctcgttc tcctcattgt
     5041 ccagaaaatc cttgtccttg atgatcttaa gcagatcatg gtaggtaccc aggcttgcgt
     5101 tgaaccgatc ttcgaccccg ctaatctcca cagagtcgaa gcattcaatt ttcttgaaat
     5161 agtcctcctt cagctgcttc acggtcacct ttctattggt cttgaacaga aggtccacaa
     5221 ttgccttctt ttgttcgcct gacaggaatg cgggcttcct catcccttcg gtgacatatt
     5281 tcaccttggt cagctcgttg tacacggtaa agtactcgta gagcagggaa tgtttcggca
     5341 ggaccttctc attggggagg ttcttatcga agttagtcat ccgttcgatg aatgactgag
     5401 cgctggctcc cttgtccacc acctcttcga agttccaggg agtgatagtt tcttctgact
     5461 ttctggtcat ccaagcaaac cggctatttc ctctggcgag gggtcccacg tagtagggga
     5521 tgcggaaagt gagaattttc tcaatcttct ccctgttgtc cttaagaaag gggtagaagt
     5581 cctcttggcg ccgaaggatg gcgtgaagct ccccaagatg gatctggtgg ggaatgctcc
     5641 cattgtcgaa agtgcgctgc ttccgcagca gatcctcccg attcagtttc acgaggagct
     5701 cttcggtgcc gtccatcttc tcaaggatag gtttgataaa cttataaaat tcctcttggg
     5761 aggcacctcc gtcgatgtag ccggcgtatc cattctttga ctggtcaaag aagatttcct
     5821 tgtacttctc cggaagctgc tgccggacga gtgccttaag cagggtcagg tcctggtggt
     5881 gctcatcata ccgcttaatc attgaagctg aaagaggggc cttggtgatt tcagtgttca
     5941 cgcggagaat gtcagacagc aggatggcgt cagaaagatt cttagcagca aggaaaaggt
     6001 cggcgtattg gtcaccgatc tgggccagga ggttgtcgag atcgtcatca taggtgtcct
     6061 tggacagttg cagcttggca tcctctgcca ggtcgaaatt agacttgaaa ttgggagtca
     6121 gcccaagtga aagagcaatc aggttcccga acagtccatt tttcttttca cccggcagct
     6181 gagcgataag attctcaagt ctgcggctct ttgacagccg ggctgacaga atggccttgg
     6241 catcgactcc gcttgcattg atggggttct cttcaaagag ctggttgtaa gtctgcacca
     6301 gctggatgaa cagtttatcc acatcgctat tgtcaggatt gaggtcgccc tcgatcagga
     6361 agtggcctct gaacttgatc atgtgggcca gggccaggta gataagccgg aggtctgcct
     6421 tgtcggttga gtccaccagc tttttgcgaa gatggtagat ggttgggtat ttctcgtgat
     6481 aggccacctc gtcgacgata ttaccgaaga tgggatgccg ttcatgcttc ttatcctctt
     6541 ccacgaggaa tgattcttcc agcctatgga agaagctgtc gtcgaccttt gccatttcgt
     6601 tggagaatat ctcttgaagg taacaaatcc ggttcttcct cctggtgtac cgccgtctag
     6661 cagtccgctt gaggcgggtt gcttcagcgg tctcccctga atcaaagagg agagctccga
     6721 tcagattttt cttgatggaa tgccggtcgg tgttcccgag caccttgaac ttcttagagg
     6781 gcaccttgta ctcgtcagtg atcacggccc acccgacgct attagtcccg atggccagtc
     6841 cgatagaata cttcttgtcc atctcagatg gttcagtgct ggtcccaggg gcagatcctt
     6901 cacttggttc ggttgacgtg ccttcttcgg tggaagtagg gcttccagct ggggaaccag
     6961 gcgcactacc ttcagatggt tctgtggagg taccagggcc tgactcgggc gttgcggatt
     7021 cgctggtgcc ttcttcagta gatgttgggg acccggcagg agaccctcca cttggtgggg
     7081 gtgcgccaga gctcggccct ccgacccaat ggttataggg ccccgcaacg tgtgtgagag
     7141 cataagggga cacggtgaca acattgtcat gggttaatgt taatgcttta tgagaaggaa
     7201 tttggttcaa ttcatttact tcagaggact gttctggacc ttcattagct gcctggtctt
     7261 tttgctctga ggccttcatt ttcttattct tagcttcttt agcctcaaac ttaatcttgt
     7321 ttagttcaaa accatgttgg ggcttattta ggtttttgtg ctggtaaaag acaagggaga
     7381 ggcgggttgg atgattacgg ttggggtgct caacaggagt ggtagcgtgc agctctcgcc
     7441 gggcacactc aatcaaaacc gagccgtgag caggtgcgat ggccacccca ccaatatttg
     7501 catccaaaaa gatgtgctca ctgtctgacc aatactcatc aatgtggggc aatttctcct
     7561 cagcaggtga cagggggtca tcagataggg gttcgtctga tggaggctca tctgcttcag
     7621 aatgctgctc atcttcttcc attggagaag aggcaaggtc ttgaggagag gtgaggaagg
     7681 agggctggtg gtttggctga tgaggcgtta gcggctcagt cacaccagtg gaaggctcag
     7741 aattaatgag gggctccatc acaggagcag acagggtggg gagaggagcc acttcgccaa
     7801 gctgtgaaat gccagggcca tcagcagctg cagcattggc accactgagt cttcccgaag
     7861 gcatcgtaca gtggggagtg ctgcttcttt ctgaaaaccc gcatgaggct gttgcgtcat
     7921 tcttcagtgg agctggtgtg gctgaagcag tcttcgggga ccaggagaag cctggagatg
     7981 cctctttcac tgggtgagga gcggatggca tcagcgaata agttttagtg ttgtctgaac
     8041 tttttaagat aaaatggggt tcggtttcac tttttacttc aggttgcacg gtctcagtgt
     8101 tactccctaa ggttggcagt gacgaaggct tactgttgtt tgttgttgtt gagttattct
     8161 tccgcttgat tcggggaata ggtttctttt ccactgccct tatcttatgt gcaagaacct
     8221 ctgtcatcat cgcagccctc ttctttccag aacggggaac aggctgagtg aaacacgttc
     8281 tttttttgcg gcggggtgcc aggacctcga tggccccaga tttgatcttg gcttccattc
     8341 cttccttgga gccaaactca tctgtgtctg aaagcttata aagaggtagc acatggagct
     8401 gctcatcttg aggaataaca cccaaagagc ggttatcttc tcgagttaag gtacaaacca
     8461 cagtgcttcc attattcatg ttgtgaatgt ccctgtgggg atgagcacag aagtccaggc
     8521 aagcagtgac cccagagaag ggtcgacctt ccttgctgcc aagccgacat tctcgggcaa
     8581 cattttcata ttccacctga ttttggtaag ctactggagc atactgctta taaattggag
     8641 ctaatcgtgt agccaaactc tgtaagttat cttcaaggtt tttttcatgt aagggagagc
     8701 ttggatcaat tctaaatctt ctggggcttg ggcttctacc aaacttacag ccattaaagt
     8761 acatactcca tgaacagcca aaagagaatg aagctccaca agtctctgga tcaattcctt
     8821 gacatgtaca ggtacgattt tcattgaggg tgcatcttct gtcggtaggg tgcccattgt
     8881 atgactttag attctctgtg agctctgtgt atagccggtc ggccattgga agagggatgc
     8941 catcccacac catgatgagc accaccatca cagcagttgg acagtggtgg cctgtacgct
     9001 gccggaccaa acaaagaact ttttcttcat cactgcttct tcttaaaacc cacttagcaa
     9061 ttggacaccc atgagagctt ttcccttctt taccggtgta cactactatt tctatcctta
     9121 ttgcgtttcc tttttgacca tacctattct ccatgatttc cctgacagca gcaacacttg
     9181 gtcctgcccc aaggtgtgta taatatgggc ctttgtcttt ttgtataact cgatcaagac
     9241 agctgcaggt gggcagggcc atggtggcgc agaattctca cgacacctga aatggaagaa
     9301 aaaaactttg aaccactgtc tgaggcttga gaatgaacca agatccaaac tcaaaaaggg
     9361 caaattccaa ggagaattac atcaagtgcc aagctggcct aacttcagtc tccacccact
     9421 cagtgtgggg aaactccatc gcataaaacc cctcccccca acctaaagac gacgtactcc
     9481 aaaagctcga gaactaatcg aggtgcctgg acggcgcccg gtactccgtg gagtcacatg
     9541 aagcgacggc tgaggacgga aaggcccttt tcctttgtgt gggtgactca cccgcccgct
     9601 ctcccgagcg ccgcgtcctc cattttgagc tccctgcagc agggccggga agcggccatc
     9661 tttccgctca cgcaactggt gccgaccggg ccagccttgc cgcccagggc ggggcgatac
     9721 acggcggcgc gaggccaggc accagagcag gccggccagc ttgagactac ccccgtccga
     9781 ttctcggtgg ccgcgctcgc aggccccgcc tcgccgaaca tgtgcgctgg gacgcacggg
     9841 ccccgtcgcc gcccgcggcc ccaaaaaccg aaataccagt gtgcagatct tggcccgcat
     9901 ttacaagact atcttgccag aaaaaaagcg tcgcagcagg tcatcaaaaa ttttaaatgg
     9961 ctagagactt atcgaaagca gcgagacagg cgcgaaggtg ccaccagatt cgcacgcggc
    10021 ggccccagcg cccaggccag gcctcaactc aagcacgagg cgaaggggct ccttaagcgc
    10081 aaggcctcga actctcccac ccacttccaa cccgaagctc gggatcaaga atcacgtact
    10141 gcagccaggt ggaagtaatt caaggcacgc aagggccata acccgtaaag aggccaggcc
    10201 cgcgggaacc acacacggca cttacctgtg ttctggcggc aaacccgttg cgaaaaagaa
    10261 cgttcacggc gactactgca cttatatacg gttctccccc accctcggga aaaaggcgga
    10321 gccagtacac gacatcactt tcccagttta ccccgcgcca ccttctctag gcaccggttc
    10381 aattgccgac ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc tctgcccact
    10441 gacgggcacc ggagccacta gtcaataatc aatgtcgacc caggtggcac ttttcgggga
    10501 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
    10561 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
    10621 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
    10681 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
    10741 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
    10801 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
    10861 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
    10921 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
    10981 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
    11041 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
    11101 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
    11161 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
    11221 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
    11281 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
    11341 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
    11401 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
    11461 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
    11521 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
    11581 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
    11641 tc
//
