LOCUS       sequence-171428-        9579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tetracycline inducible (2TO version) expression of guide RNA 
            targeted to AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9579)
  AUTHORS   Bertero A, Pawlowski M, Ortmann D, Snijders K, Yiangou L, Cardoso de
            Brito M, Brown S, Bernard WG, Cooper JD, Giacomelli E, Gambardella 
            L, Hannan NR, Iyer D, Sampaziotis F, Serrano F, Zonneveld MC, Sinha 
            S, Kotter M, Vallier L
  TITLE     Optimized inducible shRNA and CRISPR/Cas9 platforms for in vitro 
            studies of human development using hPSCs.
  JOURNAL   Development. 2016 Dec 1;143(23):4405-4418.
  PUBMED    27899508
REFERENCE   2  (bases 1 to 9579)
  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: 
            "Development."; date: "2016-12-1"; volume: "143(23)"; pages: 
            "4405-4418"
COMMENT     Sequence Label: pAAV-Puro_siKO-2TO
FEATURES             Location/Qualifiers
     source          1..9579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    295..1098
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    1105..1130
                     /label=SA
                     /note="splice acceptor site"
     CDS             1154..1207
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1217..1816
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1217..1236)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1713..1733
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1854..2078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2211..2305
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     primer_bind     2222..2241
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    2259..2277
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2287..2305
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        2340..2415
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        2494..2797
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2799..3076
                     /label=chicken beta-actin promoter
     intron          3078..4095
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4019..4041
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4103..4122
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4166..4186
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4840..4951
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    5081..5917
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(5951..5970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5952..5970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5977..5994)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5977..5993)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5986..6008)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6131..6433
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6312..6331
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             6782..7576
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6782..7576
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6836..6855)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7446..7465
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(7826..8617,8618..8686))
                     /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     8449..8468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8810..9398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9299..9318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9552..9569
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt
      301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg
      361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct
      421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc
      481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg
      541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc
      601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg
      661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt
      721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt
      781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg
      841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca
      901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat
      961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc
     1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc
     1081 tccaccccac agtggggcaa gcttctgacc tcttctcttc ctcccacagg gcctcgagag
     1141 ttctggcagc ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     1201 cggccctagg ctcgagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     1261 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     1321 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     1381 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     1441 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     1501 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     1561 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     1621 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     1681 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     1741 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     1801 caagcccggt gcctgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     1861 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1921 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1981 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2041 caatagcagg catgctgggg atgcggtggg ctctatgggt caattcgaac gctgacgtca
     2101 tcaacccgct ccaaggaatc gcgggcccag tgtcactagg cgggaacacc cagcgcgcgt
     2161 gcgccctggc aggaagatgg ctgtgaggga caggggagtg gcgccctgca atatttgcat
     2221 gtcgctatgt gttctgggaa atcaccataa acgtgaaatc cctatcagtg atagagactt
     2281 ataagttccc tatcagtgat agagatccct ggtgcaggtg gactgactca cctgcacctg
     2341 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     2401 gcaccgagtc ggtgcttttt tgtcgacctc gactagttat taatagtaat caattacggg
     2461 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2521 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2581 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2641 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     2701 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     2761 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
     2821 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
     2881 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
     2941 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
     3001 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
     3061 aagcgcgcgg cgggcgggga gtcgctgcga cgctgccttc gccccgtgcc ccgctccgcc
     3121 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
     3181 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
     3241 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
     3301 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg
     3361 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc
     3421 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa
     3481 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg
     3541 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
     3601 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggctggtg
     3661 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc
     3721 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
     3781 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct
     3841 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg
     3901 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc
     3961 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     4021 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     4081 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     4141 aaagaattaa ttcggatcca ccatgcccaa gaaaaagcgg aaggtgtccc ggctggacaa
     4201 gagcaaagtg atcaacagcg ccctggaact gctgaacgaa gtgggcatcg agggcctgac
     4261 cacccggaag ctggcccaga aactgggcgt ggaacagccc accctgtact ggcacgtgaa
     4321 gaacaagcgg gccctgctgg acgccctggc catcgagatg ctggaccggc accacacaca
     4381 cttttgcccc ctggaaggcg aaagctggca ggacttcctg cggaacaacg ccaagagctt
     4441 cagatgcgcc ctgctgagcc accgggacgg cgccaaagtg cacctgggca ccagacccac
     4501 cgagaagcag tacgagacac tggaaaacca gctggccttc ctgtgccagc agggcttcag
     4561 cctggaaaac gccctgtacg ccctgagcgc cgtgggccac tttaccctgg gctgcgtgct
     4621 ggaagatcag gaacaccagg tcgccaaaga ggaaagagag acacccacca ccgacagcat
     4681 gccccccctg ctgagacagg ccatcgagct gttcgatcat caaggcgccg agcccgcctt
     4741 cctgttcggc ctggaactga tcatctgcgg cctcgagaag cagctgaagt gcgagagcgg
     4801 ctcccccaga tgaacgcgta gctcgctgat cagcctcgac tgtgccttct agttgccagc
     4861 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4921 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4981 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg
     5041 ctgggggtct cgaggggggg cccggacagt actaagcttt actagggaca ggattggtga
     5101 cagaaaagcc ccatccttag gcctcctcct tcctagtctc ctgatattgg gtctaacccc
     5161 cacctcctgt taggcagatt ccttatctgg tgacacaccc ccatttcctg gagccatctc
     5221 tctccttgcc agaacctcta aggtttgctt acgatggagc cagagaggat cctgggaggg
     5281 agagcttggc agggggtggg agggaagggg gggatgcgtg acctgcccgg ttctcagtgg
     5341 ccaccctgcg ctaccctctc ccagaacctg agctgctctg acgcggctgt ctggtgcgtt
     5401 tcactgatcc tggtgctgca gcttccttac acttcccaag aggagaagca gtttggaaaa
     5461 acaaaatcag aataagttgg tcctgagttc taactttggc tcttcacctt tctagtcccc
     5521 aatttatatt gttcctccgt gcgtcagttt tacctgtgag ataaggccag tagccagccc
     5581 cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc ctttgtgaga
     5641 atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct actccctttc tctttctcca
     5701 tccttctttc cttaaagagt ccccagtgct atctgggaca tattcctccg cccagagcag
     5761 ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt tgagtccttg gcaagcccag
     5821 gagaggcgct caggcttccc tgtccccctt cctcgtccac catctcatgc ccctggctct
     5881 cctgcccctt ccctacaggg gttcctggct ctgctctaag ggcgaattcg cggccgctaa
     5941 attcaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     6001 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6061 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6121 atacgtacgg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     6181 ggatgtacag agtgatatta ttgacacgcc ggggcgacgg atggtgatcc ccctggccag
     6241 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     6301 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     6361 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     6421 ctggggaata taaatgtcag gcatgagatt atcaaaaagg atcttcacct agatcctttt
     6481 cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc
     6541 tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac
     6601 atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg
     6661 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     6721 aaggatctga tggcgcaggg gatcaagctc tgatcaagag acaggatgag gatcgtttcg
     6781 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     6841 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     6901 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     6961 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     7021 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     7081 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     7141 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     7201 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     7261 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     7321 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     7381 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     7441 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     7501 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     7561 tgacgagttc ttctgaatta ttaacgctta caatttcctg atgcggtatt ttctccttac
     7621 gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat gtgcgcggaa
     7681 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agattatcaa
     7741 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7801 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7861 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7921 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7981 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8041 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8101 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8161 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8221 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8281 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8341 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8401 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8461 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8521 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8581 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8641 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8701 aatattattg aagcatttat cagggttatt gtctcatgac caaaatccct taacgtgagt
     8761 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     8821 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     8881 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     8941 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     9001 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     9061 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     9121 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     9181 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     9241 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     9301 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     9361 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     9421 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     9481 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     9541 cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
