LOCUS       sequence-171440-        9481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tetracycline inducible expression of shRNA targeted to AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9481)
  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 9481)
  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_siKD
FEATURES             Location/Qualifiers
     source          1..9481
                     /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"
     primer_bind     2222..2241
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    2287..2305
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2317..2333)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2318..2334)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     enhancer        2396..2699
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2701..2978
                     /label=chicken beta-actin promoter
     intron          2980..3997
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3921..3943
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4005..4024
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4068..4088
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4742..4853
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4983..5819
                     /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(5853..5872)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5854..5872)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5879..5896)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5879..5895)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5888..5910)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6033..6335
                     /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     6214..6233
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             6684..7478
                     /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             6684..7478
                     /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(6738..6757)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7348..7367
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(7728..8519,8520..8588))
                     /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     8351..8370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8712..9300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9201..9220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9454..9471
                     /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 acgtgaaatg tctttggatt tgggaatctt
     2281 ataagttccc tatcagtgat agagatctaa gcttatcgat accgtcgacc tcgactagtt
     2341 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2401 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2461 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2521 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     2581 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     2641 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     2701 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
     2761 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
     2821 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
     2881 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
     2941 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg gagtcgctgc gacgctgcct
     3001 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     3061 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     3121 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     3181 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
     3241 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
     3301 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
     3361 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
     3421 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
     3481 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
     3541 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga
     3601 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
     3661 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
     3721 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
     3781 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     3841 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     3901 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     3961 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     4021 tgtgctgtct catcattttg gcaaagaatt aattcggatc caccatgccc aagaaaaagc
     4081 ggaaggtgtc ccggctggac aagagcaaag tgatcaacag cgccctggaa ctgctgaacg
     4141 aagtgggcat cgagggcctg accacccgga agctggccca gaaactgggc gtggaacagc
     4201 ccaccctgta ctggcacgtg aagaacaagc gggccctgct ggacgccctg gccatcgaga
     4261 tgctggaccg gcaccacaca cacttttgcc ccctggaagg cgaaagctgg caggacttcc
     4321 tgcggaacaa cgccaagagc ttcagatgcg ccctgctgag ccaccgggac ggcgccaaag
     4381 tgcacctggg caccagaccc accgagaagc agtacgagac actggaaaac cagctggcct
     4441 tcctgtgcca gcagggcttc agcctggaaa acgccctgta cgccctgagc gccgtgggcc
     4501 actttaccct gggctgcgtg ctggaagatc aggaacacca ggtcgccaaa gaggaaagag
     4561 agacacccac caccgacagc atgccccccc tgctgagaca ggccatcgag ctgttcgatc
     4621 atcaaggcgc cgagcccgcc ttcctgttcg gcctggaact gatcatctgc ggcctcgaga
     4681 agcagctgaa gtgcgagagc ggctccccca gatgaacgcg tagctcgctg atcagcctcg
     4741 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4801 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4861 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4921 tgggaagaga atagcaggca tgctgggggt ctcgaggggg ggcccggaca gtactaagct
     4981 ttactaggga caggattggt gacagaaaag ccccatcctt aggcctcctc cttcctagtc
     5041 tcctgatatt gggtctaacc cccacctcct gttaggcaga ttccttatct ggtgacacac
     5101 ccccatttcc tggagccatc tctctccttg ccagaacctc taaggtttgc ttacgatgga
     5161 gccagagagg atcctgggag ggagagcttg gcagggggtg ggagggaagg gggggatgcg
     5221 tgacctgccc ggttctcagt ggccaccctg cgctaccctc tcccagaacc tgagctgctc
     5281 tgacgcggct gtctggtgcg tttcactgat cctggtgctg cagcttcctt acacttccca
     5341 agaggagaag cagtttggaa aaacaaaatc agaataagtt ggtcctgagt tctaactttg
     5401 gctcttcacc tttctagtcc ccaatttata ttgttcctcc gtgcgtcagt tttacctgtg
     5461 agataaggcc agtagccagc cccgtcctgg cagggctgtg gtgaggaggg gggtgtccgt
     5521 gtggaaaact ccctttgtga gaatggtgcg tcctaggtgt tcaccaggtc gtggccgcct
     5581 ctactccctt tctctttctc catccttctt tccttaaaga gtccccagtg ctatctggga
     5641 catattcctc cgcccagagc agggtcccgc ttccctaagg ccctgctctg ggcttctggg
     5701 tttgagtcct tggcaagccc aggagaggcg ctcaggcttc cctgtccccc ttcctcgtcc
     5761 accatctcat gcccctggct ctcctgcccc ttccctacag gggttcctgg ctctgctcta
     5821 agggcgaatt cgcggccgct aaattcaatt cgccctatag tgagtcgtat tacaattcac
     5881 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     5941 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6001 cttcccaaca gttgcgcagc ctatacgtac ggcagtttaa ggtttacacc tataaaagag
     6061 agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg ccggggcgac
     6121 ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt
     6181 acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg
     6241 tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca
     6301 aaaacgccat taacctgatg ttctggggaa tataaatgtc aggcatgaga ttatcaaaaa
     6361 ggatcttcac ctagatcctt ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc
     6421 ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc
     6481 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
     6541 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     6601 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaagc tctgatcaag
     6661 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     6721 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     6781 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     6841 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     6901 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     6961 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     7021 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     7081 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     7141 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     7201 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     7261 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     7321 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     7381 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     7441 gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat tattaacgct tacaatttcc
     7501 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcatc aggtggcact
     7561 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7621 tatccgctca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7681 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7741 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7801 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7861 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7921 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7981 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8041 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8101 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8161 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8221 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8281 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8341 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8401 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8461 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8521 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8581 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8641 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8701 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8761 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8821 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     8881 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8941 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     9001 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9061 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     9181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     9241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     9301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     9361 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     9421 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     9481 g
//
