LOCUS       sequence-389257-       13520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral expression of hCOL4A4_C1684X in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13520)
  AUTHORS   Omachi K, Kai H, Roberge M, Miner JH
  TITLE     NanoLuc reporters identify COL4A5 nonsense mutations susceptible to 
            drug-induced stop codon readthrough.
  JOURNAL   iScience. 2022 Feb 8;25(3):103891. doi: 10.1016/j.isci.2022.103891. 
            eCollection 2022 Mar 18.
  PUBMED    35243249
REFERENCE   2  (bases 1 to 13520)
  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: "iScience.";
            date: "2022-02-8"; volume: "25(3):103891. doi"; pages: " 
            10.1016/j.isci.2022.103891. eCollection 2022 Mar 18"
COMMENT     Sequence Label: pLV-Puro_CI-hCOL4A5-Nluc_C1684X
FEATURES             Location/Qualifiers
     source          1..13520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        2211..2590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2591..2794
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2745..2765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          2930..3062
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3045..3064
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     3106..3125
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3106..3124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8228..8740
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     promoter        8756..9255
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8791..8814)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     9188..9207
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             9275..9874
                     /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(9275..9294)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9771..9791
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9888..10476
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9941..9961)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10359..10370)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10683..10916
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11045..11061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11045..11061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11058..11080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11069..11085
                     /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(11093..11099,11100..11117,11118..11123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11138..11159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11276..11293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11447..12035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11527..11546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12206..12997,12998..13066))
                     /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     12829..12848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13067..13171)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13219..13353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13256..13275)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13310..13329
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatatcgaa ttcgtgagga ccgccatgtt ggcattgatt
     2221 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2281 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2341 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2401 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2461 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2521 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2581 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
     2641 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2701 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag
     2761 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcactag
     2821 aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg cttctgacac
     2881 aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
     2941 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     3001 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     3061 aggtgtccac tcccagttca attacagctc ttaaggctag agtattaata cgactcacta
     3121 tagggctagc aaagcgatcg ccatgaaact gcgtggagtc agcctggctg ccggcttgtt
     3181 cttactggcc ctgagtcttt gggggcagcc tgcagaggct gcggcttgct atgggtgttc
     3241 tccaggatca aagtgtgact gcagtggcat aaaaggggaa aagggagaga gagggtttcc
     3301 aggtttggaa ggacacccag gattgcctgg atttccaggt ccagaagggc ctccggggcc
     3361 tcggggacaa aagggtgatg atggaattcc agggccacca ggaccaaaag gaatcagagg
     3421 tcctcctgga cttcctggat ttccagggac accaggtctt cctggaatgc caggccacga
     3481 tggggcccca ggacctcaag gtattcccgg atgcaatgga accaagggag aacgtggatt
     3541 tccaggcagt cccggttttc ctggtttaca gggtcctcca ggaccccctg ggatcccagg
     3601 tatgaagggt gaaccaggta gtataattat gtcatcactg ccaggaccaa agggtaatcc
     3661 aggatatcca ggtcctcctg gaatacaagg cctacctggt cccactggta taccagggcc
     3721 aattggtccc ccaggaccac caggtttgat gggccctcct ggtccaccag gacttccagg
     3781 acctaagggg aatatgggct taaatttcca gggacccaaa ggtgaaaaag gtgagcaagg
     3841 tcttcagggc ccacctgggc cacctgggca gatcagtgaa cagaaaagac caattgatgt
     3901 agagtttcag aaaggagatc agggacttcc tggtgaccga gggcctcctg gacctccagg
     3961 gatacgtggt cctccaggtc ccccaggtgg tgagaaaggt gagaagggtg agcaaggaga
     4021 gccaggcaaa agaggtaaac caggcaaaga tggagaaaat ggccaaccag gaattcctgg
     4081 tttgcctggt gatcctggtt accctggtga acccggaagg gatggtgaaa agggccaaaa
     4141 aggtgacact ggcccacctg gacctcctgg acttgtaatt cctagacctg ggactggtat
     4201 aactatagga gaaaaaggaa acattgggtt gcctgggttg cctggagaaa aaggagagcg
     4261 aggatttcct ggaatacagg gtccacctgg ccttcctgga cctccagggg ctgcagttat
     4321 gggtcctcct ggccctcctg gatttcctgg agaaaggggt cagaaaggtg atgaaggacc
     4381 acctggaatt tccattcctg gacctcctgg acttgacgga cagcctgggg ctcctgggct
     4441 tccagggcct cctggccctg ctggccctca cattcctcct agtgatgaga tatgtgaacc
     4501 aggccctcca ggccccccag gatctccagg tgataaagga ctccaaggag aacaaggagt
     4561 gaaaggtgac aaaggtgaca cttgcttcaa ctgcattgga actggtattt cagggcctcc
     4621 aggtcaacct ggtttgccag gtctcccagg tcctccagga tctcttggtt tccctggaca
     4681 gaaaggggaa aaaggacaag ctggtgcaac tggtcccaaa ggattaccag gcattccagg
     4741 agctccaggt gctccaggct ttcctggatc taaaggtgaa cctggtgata tcctcacttt
     4801 tccaggaatg aagggtgaca aaggagagtt gggttcccct ggagctccag ggcttcctgg
     4861 tttacctggc actcctggac aggatggatt gccagggctt cctggcccga aaggagagcc
     4921 tggtggaatt acttttaagg gtgaaagagg tccccctggg aacccaggtt taccaggcct
     4981 cccagggaat atagggccta tgggtccccc tggtttcggc cctccaggcc cagtaggtga
     5041 aaaaggcata caaggtgtgg caggaaatcc aggccagcca ggaataccag gtcctaaagg
     5101 ggatccaggt cagactataa cccagccggg gaagcctggc ttgcctggta acccaggcag
     5161 agatggtgat gtaggtcttc caggtgaccc tggacttcca gggcaaccag gcttgccagg
     5221 gatacctggt agcaaaggag aaccaggtat ccctggaatt gggcttcctg gaccacctgg
     5281 tcccaaaggc tttcctggaa ttccaggacc tccaggagca cctgggacac ctggaagaat
     5341 tggtctagaa ggccctcctg ggccacccgg ctttccagga ccaaagggtg aaccaggatt
     5401 tgcattacct gggccacctg ggccaccagg acttccaggt ttcaaaggag cacttggtcc
     5461 aaaaggtgat cgtggtttcc caggacctcc gggtcctcca ggacgcactg gcttagatgg
     5521 gctccctgga ccaaaaggtg atgttggacc aaatggacaa cctggaccaa tgggacctcc
     5581 tgggctgcca ggaataggtg ttcagggacc accaggacca ccagggattc ctgggccaat
     5641 aggtcaacct ggtttacatg gaataccagg agagaagggg gatccaggac ctcctggact
     5701 tgatgttcca ggacccccag gtgaaagagg cagtccaggg atccccggag cacctggtcc
     5761 tataggacct ccaggatcac cagggcttcc aggaaaagca ggtgcctctg gatttccagg
     5821 taccaaaggt gaaatgggta tgatgggacc tccaggccca ccaggacctt tgggaattcc
     5881 tggcaggagt ggtgtacctg gtcttaaagg tgatgatggc ttgcagggtc agccaggact
     5941 tcctggccct acaggagaaa aaggtagtaa aggagagcct ggccttccag gccctcctgg
     6001 accaatggat ccaaatcttc tgggctcaaa aggagagaag ggggaacctg gcttaccagg
     6061 tatacctgga gtttcagggc caaaaggtta tcagggtttg cctggagacc cagggcaacc
     6121 tggactgagt ggacaacctg gattaccagg accaccaggt cccaaaggta accctggtct
     6181 ccctggacag ccaggtctta taggacctcc tggacttaaa ggaaccatcg gtgatatggg
     6241 ttttccaggg cctcagggtg tggaagggcc tcctggacct tctggagttc ctggacaacc
     6301 tggctcccca ggattacctg gacagaaagg cgacaaaggt gatcctggta tttcaagcat
     6361 tggtcttcca ggtcttcctg gtccaaaggg tgagcctggt ctgcctggat acccagggaa
     6421 ccctggtatc aaaggttctg tgggagatcc tggtttgccc ggattaccag gaacccctgg
     6481 agcaaaagga caaccaggcc ttcctggatt cccaggaacc ccaggccctc ctggaccaaa
     6541 aggtattagt ggccctcctg ggaaccccgg ccttccagga gaacctggtc ctgtaggtgg
     6601 tggaggtcat cctgggcaac cagggcctcc aggcgaaaaa ggcaaacccg gtcaagatgg
     6661 tattcctgga ccagctggac agaagggtga accaggtcaa ccaggctttg gaaacccagg
     6721 accgcctgga cttccaggac tttctggcca aaagggtgat ggaggattac ctgggattcc
     6781 aggaaatcct ggccttccag gtccaaaggg cgaaccaggc tttcacggtt tccctggtgt
     6841 gcagggtccc ccaggccctc ctggttctcc gggtccagct ctggaaggac ctaaaggcaa
     6901 ccctgggccc caaggtcctc ctgggagacc aggtcctaca ggttttcaag gtctaccagg
     6961 tccagaaggt cctccaggtc tccctggaaa tggaggtatt aaaggagaga agggaaatcc
     7021 aggccaacct gggctacctg gcttgcctgg tttgaaagga gatcaaggac caccaggact
     7081 ccagggtaat cctggccggc cgggtctcaa tggaatgaaa ggagatcctg gtctccctgg
     7141 tgttccagga ttcccaggca tgaaaggacc cagtggagta cctggatcag ctggccctga
     7201 gggggaaccg ggacttattg gtcctccagg tcctcctgga ttacctggtc cttcaggaca
     7261 gagtatcata attaaaggag atgctggtcc tccaggaatc cctggccagc ctgggctaaa
     7321 gggtctacca ggaccccaag gacctcaagg cttaccaggt ccaactggcc ctccaggaga
     7381 tcctggacgc aatggactcc ctggctttga tggtgcagga gggcgcaaag gagacccagg
     7441 tctgccagga cagccaggta cccgtggttt ggatggtccc cctggtccag atggattgca
     7501 aggtccccca ggtccccctg gaacctcctc tgttgcacat ggatttctta ttacacgcca
     7561 cagccagaca acggatgcac cacaatgccc acagggaaca cttcaggtct atgaaggctt
     7621 ttctctcctg tatgtacaag gaaataaaag agcccacggt caagacttgg ggacggctgg
     7681 cagctgcctt cgtcgcttta gtaccatgcc tttcatgttc tgcaacatca ataatgtttg
     7741 caactttgct tcaagaaatg actattctta ctggctctct accccagagc ccatgccaat
     7801 gagcatgcaa cccctaaagg gccagagcat ccagccattc attagtcgat gtgcagtatg
     7861 tgaagctcca gctgtggtga tcgcagttca cagtcagacg atccagattc cccattgtcc
     7921 tcagggatgg gattctctgt ggattggtta ttccttcatg atgcatacaa gtgcaggggc
     7981 agaaggctca ggtcaagccc tagcctcccc tggttcctgc ttggaagagt ttcgttcagc
     8041 tcccttcatc gaatgtcatg ggaggggtac ctgtaactac tatgccaact cctacagctt
     8101 ttggctggca actgtagatg tgtcagacat gttcagtaaa cctcagtcag aaacgctgaa
     8161 agcaggagac ttgaggacac gaattagccg atgacaagtg tgcatgaaga ggacattctc
     8221 gagcggcgtc ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa
     8281 cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc
     8341 cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca
     8401 tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt
     8461 taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact
     8521 ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat
     8581 cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat
     8641 tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg
     8701 agtgaccggc tggcggctgt gcgaacgcat tctggcgtaa ggatccaatt ctaccgggta
     8761 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     8821 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     8881 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     8941 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     9001 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     9061 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     9121 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     9181 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     9241 tctccgggcc tttcgacctg cagttaacgc cgccatgacc gagtacaagc ccacggtgcg
     9301 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     9361 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     9421 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     9481 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     9541 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     9601 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     9661 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     9721 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     9781 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     9841 cacctggtgc atgacccgca agcccggtgc ctagacgcgt ctggaacaat caacctctgg
     9901 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     9961 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
    10021 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
    10081 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
    10141 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
    10201 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
    10261 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
    10321 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
    10381 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
    10441 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
    10501 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
    10561 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
    10621 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
    10681 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
    10741 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    10801 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    10861 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
    10921 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
    10981 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat
    11041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
    11101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
    11161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
    11221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
    11281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
    11341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
    11401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
    11461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
    11521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
    11581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
    11641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
    11701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
    11761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
    11821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
    11881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
    11941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
    12001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
    12061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
    12121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
    12181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
    12241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
    12301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
    12361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
    12421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
    12481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
    12541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
    12601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
    12661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    12721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    12781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
    12841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    12901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    12961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    13021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    13081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    13141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
    13201 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    13261 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    13321 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
    13381 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
    13441 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
    13501 tgtactacaa acttagtagt
//
