LOCUS       sequence-354945-       10899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible expression of human SOX17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10899)
  AUTHORS   Jung J, Chang Y, Jin G, Lian X, Bao X
  TITLE     Temporal Expression of Transcription Factor ID2 Improves Natural 
            Killer Cell Differentiation from Human Pluripotent Stem Cells.
  JOURNAL   ACS Synth Biol. 2022 May 24. doi: 10.1021/acssynbio.2c00017.
  PUBMED    35608547
REFERENCE   2  (bases 1 to 10899)
  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: "ACS Synth 
            Biol. 2022 May 24. doi: 10.1021/acssynbio.2c00017."
COMMENT     Sequence Label: AAVS1-Puro Xlone SOX17
FEATURES             Location/Qualifiers
     source          1..10899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    77..913
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    987..1008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1023..1028,1029..1046,1047..1053)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1061..1077
                     /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     1085..1101
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1444..1463
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1462..1517
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1563..1582
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1692..2168
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1950..1969
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(2212..2958)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(2959..2982)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2982..2991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3011..3222)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        3264..3631
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    3272..3290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3308..3326
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3344..3362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3380..3398
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3416..3434
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3452..3470
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3488..3506
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             4592..4609
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             4598..4615
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     polyA_signal    5069..5203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5147..5166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    5228..5452
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5441..5458
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5490..6089)
                     /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(5573..5593)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6070..6089
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(6099..6152)
                     /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"
     misc_feature    6176..6201
                     /label=SA
                     /note="splice acceptor site"
     misc_feature    6208..7011
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(7050..7068)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7089..7105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7113..7129
                     /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(7137..7143,7144..7161,7162..7167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7182..7203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7320..7337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7491..8079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7571..7590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(8203..8271,8272..9063)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8421..8440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     complement(9424..9443)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10038..10057
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(10460..10762)
                     /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     complement(10562..10581)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     join(10885..10899,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(10899,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg aatttagcgg
       61 ccgcgaattc gcccttagag cagagccagg aacccctgta gggaaggggc aggagagcca
      121 ggggcatgag atggtggacg aggaaggggg acagggaagc ctgagcgcct ctcctgggct
      181 tgccaaggac tcaaacccag aagcccagag cagggcctta gggaagcggg accctgctct
      241 gggcggagga atatgtccca gatagcactg gggactcttt aaggaaagaa ggatggagaa
      301 agagaaaggg agtagaggcg gccacgacct ggtgaacacc taggacgcac cattctcaca
      361 aagggagttt tccacacgga cacccccctc ctcaccacag ccctgccagg acggggctgg
      421 ctactggcct tatctcacag gtaaaactga cgcacggagg aacaatataa attggggact
      481 agaaaggtga agagccaaag ttagaactca ggaccaactt attctgattt tgtttttcca
      541 aactgcttct cctcttggga agtgtaagga agctgcagca ccaggatcag tgaaacgcac
      601 cagacagccg cgtcagagca gctcaggttc tgggagaggg tagcgcaggg tggccactga
      661 gaaccgggca ggtcacgcat cccccccttc cctcccaccc cctgccaagc tctccctccc
      721 aggatcctct ctggctccat cgtaagcaaa ccttagaggt tctggcaagg agagagatgg
      781 ctccaggaaa tgggggtgtg tcaccagata aggaatctgc ctaacaggag gtgggggtta
      841 gacccaatat caggagacta ggaaggagga ggcctaagga tggggctttt ctgtcaccaa
      901 tcctgtccct agtaaagctt agtactgtcg atccgctggc acgacaggtt tcccgactgg
      961 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1021 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1081 cacacaggaa acagctatga ccatgattac gccaagcttg ggctgcaggt cgagggatct
     1141 ccataagaga agagggacag ctatgactgg gagtagtcag gagaggagga aaaatctggc
     1201 tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca caaaacaaaa
     1261 tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat ggacagcagg
     1321 gggctgtttc atatactgat gacctcttta tagccaacct ttgttcatgg cagccagcat
     1381 atgggcatat gttgccaaac tctaaaccaa atactcattc tgatgtttta aatgatttgc
     1441 cctcccatat gtccttccga gtgagagaca caaaaaattc caacacacta ttgcaatgaa
     1501 aataaatttc ctttattagc cagaagtcag atgctcaagg ggcttcatga tgtccccata
     1561 atttttggca gagggaaaaa gatctcagtg gtatttgtga gccagggcat tggccacacc
     1621 agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattcacgcg cctggttacc
     1681 tacaaaatca gaaggacagg gaagggagca gtggttcacg cctgtaatcc cagcaatttg
     1741 ggaggccaag gtgggtagat cacctgagat taggagttgg agaccagcct ggccaatatg
     1801 gtgaaacccc gtctctacca aaaaaacaaa aattagctga gcctggtcat gcatgcctgg
     1861 aatcccaaca actcgggagg ctgaggcagg agaatcgctt gaacccagga ggcggagatt
     1921 gcagtgagcc aagattgtgc cactgcactc cagcttggtt cccaatagac cccgcaggcc
     1981 ctacaggttg tcttcccaac ttgccccttg ctccatacca cccccctcca ccccataata
     2041 ttatagaagg acacctagtc agacaaaatg atgcaactta attttattag gacaaggctg
     2101 gtgggcactg gagtggcaac ttccagggcc aggagaggca ctggggaggg gtcacaggga
     2161 tgccacccgt agatctctcg agcagcgctc gatgatatcg aattcacgcg tttacccggg
     2221 gagcatgtca aggtcaaaat cgtcaagagc gtcagcaggc agcatatcaa ggtcaaagtc
     2281 gtcaagggca tcggctggga gcatgtctaa gtcaaaatcg tcaagggcgt cggtcggccc
     2341 gccgctttcg cactttagct gtttctccag gccacatatg attagttcca ggccgaaaag
     2401 gaaggcaggt tcggctccct gccggtcgaa cagctcaatt gcttgtttca gaagtggggg
     2461 catagaatcg gtggtaggtg tctctctttc ctcttttgct acttgatgct cctgttcctc
     2521 caatacgcag cccagtgtaa agtggcccac ggcggacaga gcgtacagtg cgttctccag
     2581 ggagaagcct tgctgacaca ggaacgcgag ctgattttcc agggtttcgt actgtttctc
     2641 tgttgggcgg gtgccgagat gcactttagc cccgtcgcga tgtgagagga gagcacagcg
     2701 gtatgacttg gcgttgttcc gcagaaagtc ttgccatgac tcgccttcca gggggcagga
     2761 gtgggtatga tgcctgtcca gcatctcgat tggcagggca tcgagcaggg cccgcttgtt
     2821 cttcacgtgc cagtacaggg taggctgctc aactcccagc ttttgagcga gtttccttgt
     2881 cgtcaggcct tcgataccga ctccattgag taattccaga gcagagttta tgactttgct
     2941 cttgtccagt ctagacatct tatcgtcatc gtctttgtaa tccatggtgg cggatcccgc
     3001 gtcacgacac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
     3061 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
     3121 cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg ccgacccctc
     3181 cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc
     3241 cactcgagtg gaattatcac ctcgagttta ctccctatca gtgatagaga acgtatgaag
     3301 agtttactcc ctatcagtga tagagaacgt atgcagactt tactccctat cagtgataga
     3361 gaacgtataa ggagtttact ccctatcagt gatagagaac gtatgaccag tttactccct
     3421 atcagtgata gagaacgtat ctacagttta ctccctatca gtgatagaga acgtatatcc
     3481 agtttactcc ctatcagtga tagagaacgt ataagctttg cttatgtaaa ccagggcgcc
     3541 tataaaagag tgctgatttt ttgagtaaac ttcaattcca caacactttt gtcttatacc
     3601 aactttccgt accacttcct accctcgtaa aggtaccgcc accatgagca gcccggatgc
     3661 gggatacgcc agtgacgacc agagccagac ccagagcgcg ctgcccgcgg tgatggccgg
     3721 gctgggcccc tgcccctggg ccgagtcgct gagccccatc ggggacatga aggtgaaggg
     3781 cgaggcgccg gcgaacagcg gagcaccggc cggggccgcg ggccgagcca agggcgagtc
     3841 ccgtatccgg cggccgatga acgctttcat ggtgtgggct aaggacgagc gcaagcggct
     3901 ggcgcagcag aatccagacc tgcacaacgc cgagttgagc aagatgctgg gcaagtcgtg
     3961 gaaggcgctg acgctggcgg agaagcggcc cttcgtggag gaggcagagc ggctgcgcgt
     4021 gcagcacatg caggaccacc ccaactacaa gtaccggccg cggcggcgca agcaggtgaa
     4081 gcggctgaag cgggtggagg gcggcttcct gcacggcctg gctgagccgc aggcggccgc
     4141 gctgggcccc gagggcggcc gcgtggccat ggacggcctg ggcctccagt tccccgagca
     4201 gggcttcccc gccggcccgc cgctgctgcc tccgcacatg ggcggccact accgcgactg
     4261 ccagagtctg ggcgcgcctc cgctcgacgg ctacccgttg cccacgcccg acacgtcccc
     4321 gctggacggc gtggaccccg acccggcttt cttcgccgcc ccgatgcccg gggactgccc
     4381 ggcggccggc acctacagct acgcgcaggt ctcggactac gctggccccc cggagcctcc
     4441 cgccggtccc atgcaccccc gactcggccc agagcccgcg ggtccctcga ttccgggcct
     4501 cctggcgcca cccagcgccc ttcacgtgta ctacggcgcg atgggctcgc ccggggcggg
     4561 cggcgggcgc ggcttccaga tgcagccgca acaccagcac cagcaccagc accagcacca
     4621 ccccccgggc cccggacagc cgtcgccccc tccggaggca ctgccctgcc gggacggcac
     4681 ggaccccagt cagcccgccg agctcctcgg ggaggtggac cgcacggaat ttgaacagta
     4741 tctgcacttc gtgtgcaagc ctgagatggg cctcccctac caggggcatg actccggtgt
     4801 gaatctcccc gacagccacg gggccatttc ctcggtggtg tccgacgcca gctccgcggt
     4861 atattactgc aactatcctg acgtgtagac tagtagacca cctcccctgc gagctaagct
     4921 ggacagccaa tgacgggtaa gagagtgaca tttttcacta acctaagaca ggagggccgt
     4981 cagagctact gcctaatcca aagacgggta aaagtgataa aaatgtatca ctccaaccta
     5041 agacaggcgc agcttccgag ggatttgaga tccagacatg ataagataca ttgatgagtt
     5101 tggacaaacc aaaactagaa tgcagtgaaa aaaatgcctt atttgtgaaa tttgtgatgc
     5161 tattgcctta tttgtaacca ttataagctg caataaacaa gttctagtca ataatcaatg
     5221 tcgagaccca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
     5281 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     5341 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     5401 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     5461 gatcagcggg tttaaacggg ccctctagat caggcaccgg gcttgcgggt catgcaccag
     5521 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     5581 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
     5641 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     5701 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     5761 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     5821 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     5881 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     5941 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     6001 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     6061 aggcgcaccg tgggcttgta ctcggtcatc tcgagcctag ggccgggatt ctcctccacg
     6121 tcaccgcatg ttagaagact tcctctgccc tctccgctgc cagatctctc gaggccctgt
     6181 gggaggaaga gaagaggtca gaagcttgcc ccactgtggg gtggagggga cagataaaag
     6241 tacccagaac cagagccaca ttaaccggcc ctgggaatat aaggtggtcc cagctcgggg
     6301 acacaggatc cctggaggca gcaaacatgc tgtcctgaag tggacatagg ggcccgggtt
     6361 ggaggaagaa gactagctga gctctcggac ccctggaaga tgccatgaca gggggctgga
     6421 agagctagca cagactagag aggtaagggg ggtaggggag ctgcccaaat gaaaggagtg
     6481 agaggtgacc cgaatccaca ggagaacggg gtgtccaggc aaagaaagca agaggatgga
     6541 gaggtggcta aagccaggga gacggggtac tttggggttg tccagaaaaa cggtgatgat
     6601 gcaggcctac aagaagggga ggcgggacgc aagggagaca tccgtcggag aaggccatcc
     6661 taagaaacga gagatggcac aggccccaga aggagaagga aaagggaacc cagcgagtga
     6721 agacggcatg gggttgggtg agggaggaga gatgcccgga gaggacccag acacggggag
     6781 gatccgctca gaggacatca cgtggtgcag cgccgagaag gaagtgctcc ggaaagagca
     6841 tccttgggca gcaacacagc agagagcaag gggaagaggg agtggaggaa gacggaacct
     6901 gaaggaggcg gcagggaagg atctgggcca gccgtagagg tgacccaggc cacaagctgc
     6961 agacagaaag cggcacaggc ccaggggaga gaatgctggt cagagaaagc aaagggcgaa
     7021 ttcgtttaaa cctgcaggac tagctagtcc ctttagtgag ggttaattct gagcttggcg
     7081 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7141 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7201 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7261 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7321 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7381 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7441 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7501 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7561 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7621 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7681 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7741 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7801 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7861 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7921 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7981 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     8041 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     8101 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagacaa
     8161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     8221 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     8281 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     8341 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     8401 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     8461 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc
     8521 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     8581 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     8641 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     8701 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     8761 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     8821 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8881 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     8941 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     9001 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     9061 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     9121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgagcggata catatttgaa
     9181 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     9241 gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     9301 agcgttaata attcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     9361 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     9421 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     9481 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     9541 gccatgcatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     9601 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     9661 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     9721 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     9781 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     9841 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     9901 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     9961 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
    10021 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
    10081 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
    10141 cagagcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
    10201 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
    10261 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
    10321 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
    10381 agcaccgttt ctgcggactg gctttctacg tgaaaaggat ctaggtgaag atcctttttg
    10441 ataatctcat gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt
    10501 cattttcgcg gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg
    10561 ccatatcggt ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt
    10621 cacgggagac tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc
    10681 ccggcgtgtc aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt
    10741 tataggtgta aaccttaaac tgccgtacgt ataggctgcg caactgttgg gaagggcgat
    10801 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
    10861 taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
