LOCUS       sequence-437285-       12592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus expressing mutant tRNA synthetase for incorporation of 
            noncanonical amino acids in nascent proteins plus mCherry reporter 
            and puro resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12592)
  AUTHORS   Sridharan D, Dougherty JA, Ahmed U, Sanghvi SK, Alvi SB, Park KH, 
            Islam H, Knoblaugh SE, Singh H, Kirby ED, Khan M
  TITLE     Bioorthogonal non-canonical amino acid tagging to track transplanted
            human induced pluripotent stem cell-specific proteome.
  JOURNAL   Stem Cell Res Ther. 2024 Jun 26;15(1):186. doi: 
            10.1186/s13287-024-03792-3.
  PUBMED    38926849
REFERENCE   2  (bases 1 to 12592)
  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 
            Res Ther."; date: "2024-06-26"; volume: "15(1):186. doi"; pages: " 
            10.1186/s13287-024-03792-3"
COMMENT     Sequence Label: EF1a-L274GMmMetRS-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..12592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(104..123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      917..1505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1406..1425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1659..1676
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1793..1814
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1829..1834,1835..1852,1853..1859)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1867..1883
                     /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     1872..1894
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1891..1907
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1891..1907
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1926..1946
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1928..1946
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1974..2200
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2201..2381
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2428..2553
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3046..3279
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3464..3508
                     /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"
     primer_bind     3806..3826
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        3985..5163
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4216..5154
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5111..5131
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        5180..5198
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5219..5228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5228..5251
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7316..7327
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             complement(7568..7579)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             7958..8011
                     /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             8012..8719
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8157..8175)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8425..8444
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             8720..8749
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    8772..9360
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8825..8845)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9243..9254)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    9393..9617
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    9659..9776
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        9825..10335
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     10254..10272
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             10357..10956
                     /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(10357..10376)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10853..10873
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             11084..11317
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    11389..11510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11426..11445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11480..11499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      11550..11685
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(11706..11724)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11734..11751)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11734..11750)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11743..11765)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11892..12347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11979..11998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12189..12210
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        12373..12477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12478..12546,12547..12592,1..746)
                     /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"
ORIGIN
        1 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
       61 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      601 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
      781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
      841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
      901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1561 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1621 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1681 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1741 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     1801 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1861 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     1921 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
     1981 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
     2041 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
     2101 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
     2161 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
     2221 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     2281 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     2341 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     2401 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
     2461 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
     2521 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
     2581 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
     2641 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     2701 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     2761 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     2821 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     2881 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     2941 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     3001 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     3061 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     3121 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     3181 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     3241 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     3301 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     3361 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     3421 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     3481 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     3541 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     3601 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     3661 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     3721 agacagagac agatccattc gattagtgaa cggatctcga cggtatcgat cacgagacta
     3781 gcctcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca
     3841 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
     3901 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     3961 attatgtttt aaaatggact atcaggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     4021 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     4081 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     4141 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     4201 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     4261 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     4321 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
     4381 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     4441 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     4501 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
     4561 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     4621 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     4681 gctggccggc ctgctctggt gcctggtctc gcgccgccgt gtatcgcccc gccctgggcg
     4741 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     4801 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     4861 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     4921 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
     4981 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     5041 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     5101 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     5161 tgaggaatta gcttggtact aatacgactc actataggga gacccaagct ggctagccgc
     5221 caccatggat tacaaggatg atgatgataa gatgagactg ttcgtgagcg agggttcccc
     5281 ggggagcctg cccgtgctgg ctgcggccgc gagggcccgg ggtcgggcgg agctgctcat
     5341 cagcaccgta ggccccgaag agtgtgtggt accattcctt acccggccta aggtccctgt
     5401 cttgcagctg gatagtggca actacctctt ctctgctagt gcaatctgcc gatatttttt
     5461 tctgttatgt ggctgggaac aagatgatct caccaaccag tggctggaat gggaggcaac
     5521 agaactgcag ccagttctgt ctgctgccct acactgtcta gtggttcaag gcaagaaagg
     5581 ggaagatata cttggcccac ttcggagagt cctgactcac attgatcaca gcttgagtcg
     5641 tcaaaactgt cctttcctgg ctggggacac agaatctcta gctgacattg ttttgtgggg
     5701 agcactgtat cctttactgc aagacccagc ttacctccct gaggagttgg gtgccctgca
     5761 aagttggttc cagacactga gtacccagga accgtgtcag cgagctgcag agacggtgct
     5821 aaaacagcag ggtgtcctgg cacttcgtct gtacctccag aaacagccac agcctcagcc
     5881 cccgcctcct gaggggagaa ctgtcagcaa cgagctggag gaagaggaac tggctacctt
     5941 gtctgaggag gacatcgtta cagctgttgc cgcgtgggag aagggtctgg aaagcctgcc
     6001 tccgctaaag ctccagcagc atccagtgtt gcctgtgcct ggagagagga atgttctcat
     6061 caccagtgcc ggaccctatg tcaacaatgt cccccacctt ggaaacatca ttggctgtgt
     6121 gctcagtgct gatgtctttg caaggtattg tcgccttcgc cagtggaata ccctctatct
     6181 gtgtggtaca gatgagtatg gtactgcgac agagaccaag gccatggagg agggcctaac
     6241 cccacgggaa atctgtgaca agtaccatgc catccatgct gacatctacc gctggttcgg
     6301 catatcgttc gatactttcg ggcgcactac cactcctcag cagaccaaaa tcacccagga
     6361 catcttccag aggttgctga cccgggggtt tgtgctgcga gatactgtgg agcagcttcg
     6421 gtgtgagcgg tgtgcacgtt tcctggctga ccgctttgtg gagggtgtgt gtcccttctg
     6481 tggctatgaa gaggcccgag gtgaccagtg tgacaggtgt ggcaagctca tcaatgccat
     6541 tgagctcaag aaaccacagt gcaaaatctg ccgctcctgc cctgtggtga ggtcctcaca
     6601 gcacctgttt ctagacttgc ctaagttgga aaagcgtctg gaggactggt tggggaagac
     6661 agtgcctggc agtgactgga cacccaatgc caggttcatt atacgttcct ggcttcgaga
     6721 tggcctcaag ccacgatgca tcaccagaga cctcaaatgg ggaacgcctg tgcccttgga
     6781 aggttttgag gacaaggtat tttacgtctg gtttgatgct actattggct acgtgtccat
     6841 cacagccaac tacacagacc aatgggagaa atggtggaag aacccagaac aagtggacct
     6901 ttaccagttc atggccaaag acaatgttcc cttccatggc ttggtctttc cgtgttcagt
     6961 cctaggagct gaggacaact acaccctggt caagcacatc attgctacag agtacctgaa
     7021 ctatgaggat gggaaattct ctaagagccg gggcatagga gtgtttggag acatggccaa
     7081 ggatacagga atccctgctg acatctggcg attctatctg ctatacattc ggcctgaggg
     7141 ccaggacagt gccttctcct ggacagactt gttgattaaa aacaattctg agctgctcaa
     7201 caacctgggc aacttcatca acagagctgg catgtttgtt tctaagtttt ttggcggttg
     7261 tgtgcctgag atggcgctaa cccctgatga cagacgcctg gtggcccatg tctcttggga
     7321 actccagcac tatcaccagc tgttggaaaa ggttcggatc cgggatgcct tgcgcagtat
     7381 cctcaccata tctcgccatg gcaaccaata cattcaagtg aatgagccct ggaaacggat
     7441 taaaggtggt gagatggaca ggcagcgggc aggcacagtg acaggcatgg cagtgaacat
     7501 ggctgccttg ctgtctgtca tgctgcagcc atacatgccc acagtcagct ctaccatcca
     7561 gacccagctg cagctcccac ctgcagcctg ccgcatcctt gccacaagct tcatttgtac
     7621 cttgccagca ggccaccgaa ttggcacagt cagtcctttg ttccaaaaac tggaaaatga
     7681 ccagattgaa aatttgaggc agcgctttgg agggggtcag gctaaaggct cccccaagcc
     7741 agcagctgtg gaggcagtta cagcagcagg ctcgcagcac atacaaacgc tgacggatga
     7801 ggtgaccaag cagggcaacg tcgtccggga actgaaagca cagaaggcag acaagaacca
     7861 ggttgctgca gaggtggcta aactcttgga tctaaagaaa cagttggctt tggctgaggg
     7921 gaaacccatt gaaactccta aaggcaagaa gaaaaaggaa gggagaggaa gcctcctaac
     7981 atgcggtgac gtggaggaga acccaggacc aatggtgagc aagggcgagg aggataacat
     8041 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     8101 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     8161 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     8221 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     8281 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     8341 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     8401 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     8461 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     8521 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     8581 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     8641 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     8701 catggacgag ctgtacaagg aacaaaaact tatttctgaa gaagatctgt aacgatatca
     8761 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     8821 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     8881 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     8941 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     9001 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     9061 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     9121 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     9181 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     9241 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     9301 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     9361 atcgataccg agcgctgctc gagagatctc gactgtgcct tctagttgcc agccatctgt
     9421 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     9481 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     9541 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     9601 tgcggtgggc tctatgggaa ttctcgacct cgagacaaat ggcagtattc atccacaatt
     9661 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     9721 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     9781 tttattacag ggacagcaga gatccacttt ggccgcggct cgagggggtt ggggttgcgc
     9841 cttttccaag gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg
     9901 aaacgcagcg gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc
     9961 cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt
    10021 cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc
    10081 actagtaccc tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg
    10141 gctgtggcca atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg
    10201 gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc
    10261 gcattctgca agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc
    10321 gacctctctc cccaggggga tccaccggag cttaccatga ccgagtacaa gcccacggtg
    10381 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
    10441 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
    10501 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
    10561 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
    10621 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
    10681 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
    10741 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
    10801 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
    10861 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
    10921 cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga cccgcagcgc
    10981 ccgaccgaaa ggagcgcacg accccatgca tcggtacctt taagaccaat gacttacaag
    11041 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
    11101 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
    11161 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
    11221 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
    11281 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
    11341 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
    11401 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
    11461 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
    11521 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
    11581 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
    11641 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
    11701 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
    11761 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
    11821 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
    11881 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
    11941 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
    12001 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
    12061 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
    12121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
    12181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
    12241 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
    12301 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
    12361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
    12421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
    12481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
    12541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gg
//
