LOCUS       sequence-409502-       16522 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector for overexpressing transcription factor ORFs with 
            unique 24-bp barcodes. Barcodes facilitate identification of 
            transcription factors in pooled screens..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16522)
  AUTHORS   Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine 
            VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh
            OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F
  TITLE     A transcription factor atlas of directed differentiation.
  JOURNAL   Cell. 2023 Jan 5;186(1):209-229.e26. doi: 
            10.1016/j.cell.2022.11.026.
  PUBMED    36608654
REFERENCE   2  (bases 1 to 16522)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5";
            volume: "186"; issue: "1"; pages: "209-229.e26"
COMMENT     Sequence Label: TFORF0751
FEATURES             Location/Qualifiers
     source          1..16522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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"
     promoter        1770..1980
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        1792..1988
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1839..1974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2020..2622
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(2020..2039)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2516..2536
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2665..2782
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2920..4098
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3151..4089
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4046..4066
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    10664..10675
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     misc_feature    12379..12967
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12432..12452)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12850..12861)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             13039..13272
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    13350..13471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13387..13406)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13441..13460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      13511..13646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(13666..13685)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(13667..13685)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13695..13712)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13695..13711)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13704..13726)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      13853..14308
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13940..13959)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14150..14171
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        14334..14438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14439..14507,14508..15299)
                     /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(14657..14676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      15470..16058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     15959..15978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     16212..16229
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    16346..16367
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(16382..16387,16388..16405,16406..16412)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    16420..16436
                     /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     16425..16447
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     16444..16460
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16444..16460
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     16479..16499
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        16481..16499
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa
     1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca
     1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1981 tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta caagcccacg
     2041 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     2101 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     2161 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     2221 gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     2281 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     2341 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     2401 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     2461 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     2521 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     2581 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct agatcttgag
     2641 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     2701 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     2761 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg
     2821 ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     2881 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     2941 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
     3001 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     3061 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     3121 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
     3181 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
     3241 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     3301 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     3361 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     3421 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     3481 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     3541 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     3601 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     3661 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     3721 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     3781 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     3841 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     3901 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     3961 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     4021 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     4081 ccatttcagg tgtcgtgagg ctagcaccgc caccatgcct cgaactaaac aaattcatcc
     4141 cagaaatcta agagacaaaa ttgaagaagc acaaaaagaa cttaatgggg cagaagtttc
     4201 aaaaaaagaa atcttacagg ctggtgttaa aggaacttcg gaatccctta aaggtgtgaa
     4261 acgcaaaaag atcgtagctg agaatcacct gaaaaaaata ccaaaatccc cactgagaaa
     4321 tcctcttcag gcaaaacata aacaaaatac agaagagtca tctttcgccg ttcttcatag
     4381 tgcttcggag tctcacaaga aacagaatta tattcctgta aaaaatggga agcagtttac
     4441 caaacaaaat ggagaaacac ctggaataat tgctgaagcc tcaaaatctg aagaatctgt
     4501 ctccccaaag aagcccttgt ttctgcagca accatctgaa ctgcgtagat ggagatccga
     4561 aggcgctgat cctgccaaat tcagtgacct cgatgaacaa tgtgactcaa gttccttgtc
     4621 aagtaaaacc aggactgaca atagcgaatg catctcttct cattgtggca ctacgtcccc
     4681 ctcctataca aacactgcat tcgatgtctt actgaaagca atggagccag aactgagcac
     4741 cttgtcacaa aagggctcac cttgtgcaat taagacagaa aaactgaggc caaataaaac
     4801 tgcacgttcc cctcccaaat taaaaaacag ttcaatggat gccccaaatc agacttcaca
     4861 ggaattggtt gctgaatcac agtcttcttg tacctcatac acagtccata tgtctgctgc
     4921 tcagaagaat gagcaagggg caatgcagtc agcttctcat ttgtatcatc aacatgaaca
     4981 ctttgttccc aaatccaacc aacataatca acagcttccg gggtgttcag gtttcacagg
     5041 atcactgaca aatctgcaaa atcaagagaa tgccaaactt gaacaggttt ataatatagc
     5101 agtgacatca tctgtaggcc taacttcacc ttccagtaga tctcaggtta ctcctcaaaa
     5161 ccagcaaatg gattctgctt cacctttgtc aataagtccg gctaattcta cacagtcgcc
     5221 ccccatgcca atctataatt caactcatgt tgcctctgtt gttaatcaaa gcgtagagca
     5281 aatgtgcaat cttcttctga aagatcagaa gccaaaaaaa caaggaaaat atatttgtga
     5341 gtattgcaat agagcatgtg caaagcctag tgtgctttta aagcatatcc gctcccacac
     5401 tggagagcga ccctatccct gtgtgacttg tggattttca tttaagacta aaagtaatct
     5461 gtataagcac aagaaatccc acgcacatac tatcaaactg ggtcttgtct tgcaaccaga
     5521 tgctggtggc ttgttcttgt cccacgagtc ccccaaagca cttagtattc attcagacgt
     5581 agaagacagt ggggagagcg aggaggaagg cgccactgat gagagacagc atgacctggg
     5641 cgccatggag ctgcagcctg tgcacataat aaagaggatg tcaaatgctg aaactttact
     5701 aaaatcaagc ttcactccaa gcagtccaga aaatgtgata ggtgactttt tgctacagga
     5761 cagatctgca gaatcacaag ctgtgacaga gttaccgaaa gttgtggtcc accatgtcac
     5821 tgtgtccccc ttaagaactg acagtccaaa ggccatggat cccaagcctg aactttctag
     5881 tgcacaaaag cagaaggacc ttcaggtgac aaacgtacag ccactttcag ccaacatgtc
     5941 ccagggtgga gtctccaggt tggagactaa tgagaattcc caccagaaag gcgacatgaa
     6001 tccactggaa ggaaagcaag actctcacgt aggaacggta cacgcccagc tacaaaggca
     6061 gcaggctacc gattactccc aagagcagca aggaaagctc ctgagtcctc gaagtttagg
     6121 aagtacggat tctggttact tttcacgttc tgaaagtgcc gatcaaacag tgagtccacc
     6181 aactcccttt gccagaaggt tacccagcac agaacaagac tctggaagga gtaacggacc
     6241 ctctgcagct cttgtcacca cgtcaacacc ctctgctttg cccacagggg aaaaggcatt
     6301 gcttttacca ggtcagatgc gcccaccttt ggccacaaaa acacttgagg agcggatatc
     6361 gaagcttatc tcagacaatg aagctttggt agatgacaag caactggata gtgtgaagcc
     6421 gcggagaacc tcactgtcaa gacgaggaag cattgattcc cccaaatcat acatatttaa
     6481 agattctttc cagtttgatt taaaaccagt gggacggaga acaagttcaa gctctgatat
     6541 accgaagtca cctttcaccc ctactgaaaa atcaaagcaa gtgtttcttc tgtctgtacc
     6601 ttcacttgac tgtttaccta tcacaagaag taattccatg ccgaccacag gttattcagc
     6661 agtacctgca aatataatac ctcctcctca tccactaaga ggaagtcagt catttgatga
     6721 caaaattggc gctttctatg atgatgtctt tgtatcggga cctaacgctc ctgtgcccca
     6781 gagtgggcat ccccgtacac ttgtgagaca agcagccata gaagactctt cagcaaatga
     6841 aagtcatgtt cttggtactg gacagtccct ggatgagagc caccaaggat gccatgctgc
     6901 tggtgaagcc atgtcagtga ggagcaaggc actggcacaa ggcccacata tagaaaaaaa
     6961 gaagtctcat caagggcgag ggacaatgtt tgagtgtgaa acttgtagaa acaggtatag
     7021 gaaactggaa aattttgaaa atcataagaa attttactgt tctgagttac atggaccaaa
     7081 aacaaaggta gccatgagag aacctgagca cagccctgtg cccggcggtc tgcagcctca
     7141 gattctacac tacagagtcg ctgggtcctc cggcatctgg gaacagacgc cccagataag
     7201 aaaaaggagg aaaatgaaaa gtgttgggga tgatgaagaa cttcagcaaa atgaaagtgg
     7261 aacatctcca aaaagttctg aaggccttca gtttcagaat gctctgggct gtaatcccag
     7321 tttgcctaaa cataatgtta ccataagaag tgaccagcag cataaaaata tacagttgca
     7381 aaactcccat attcaccttg ttgccagggg ccctgagcag accatggatc ccaagctgtc
     7441 gaccatcatg gaacaacaga taagttcagc agcccaggac aagatagaac tgcagagaca
     7501 cggaactgga atctctgtca tccagcacac caactccctg agcaggccca actcatttga
     7561 caagcctgag ccttttgaaa gagcctcccc agtttctttc caggagctga atagaacggg
     7621 gaagtccggg tctctaaaag tgataggaat ctcccaagag gaaagtcacc cttctcggga
     7681 cgggtctcat cctcaccagc ttgcactatc agacgctctc agaggagaac ttcaggaaag
     7741 ctccagaaag agtccaagtg aacgacatgt gttaggacag ccctcaagac ttgtccggca
     7801 gcacaacatc caagttccag agattttggt cacagaagaa ccagatcgag acctggaagc
     7861 tcaatgccat gatcaagaaa agtcagagaa gttcagttgg ccccagcgta gtgaaacctt
     7921 gtcaaaattg ccaacagaga aactgccacc caaaaagaaa aggctccgtc tggctgagat
     7981 agaacattcc tcaacagaat cgagctttga ttccactctc tccaggagtc taagtaggga
     8041 gagcagttta tctcacactt caagtttctc agcctcttta gacatagagg acgtttctaa
     8101 aacggaggct tcccccaaaa tcgattttct aaataaagcc gagtttctta tgattccagc
     8161 tggcttgaat actctgaatg ttcctggatg tcaccgggaa atgaggcgta ctgcatcaga
     8221 acagattaat tgcacgcaaa cgtcaatgga ggtctctgat ctcagaagca aatcattcga
     8281 ttgtggaagc atcaccccac cccagacaac accacttact gaattgcagc ctccatcttc
     8341 accttctcga gtgggagtga ctgggcatgt gcctctctta gaaagaagga gaggcccact
     8401 ggtacggcaa atatctttaa acatagcccc agatagtcat ctgtctcctg tacacccaac
     8461 atctttccaa aatactgctc ttcccagtgt gaatgcagtg ccatatcagg ggcctcagct
     8521 cactagtaca tctttagctg agttttctgc aaatactttg cactctcaga ctcaggttaa
     8581 ggatctgcag gcagaaacat caaactccag ctctaccaac gtttttcctg ttcaacagct
     8641 ctgtgatatc aatttgttaa atcaaatcca tgcaccgcct agccaccaga gcacacagct
     8701 atctctgcaa gtgtctacgc agggtagcaa gccagataaa aattctgttt tatctgggtc
     8761 ttctaaaagt gaggattgct ttgctcccaa ataccaattg cattgtcagg ttttcacttc
     8821 aggcccatct tgctcttcta atcctgtgca ttctttgcca aatcaagtta tttcagatcc
     8881 agttggaaca gatcattgtg tgacatcagc aacattacca accaaattaa ttgacagcat
     8941 gtctaattcg catcctctgc taccaccaga gctcaggccc cttggaagtc aggtgcagaa
     9001 ggtgccatca tcattcatgc tgcccatacg cctgcagagt agtgttcctg cttactgttt
     9061 tgctacactc acatccctgc cacaaatact agtgacccaa gatctgccca atcagccaat
     9121 ttgccagact aatcatagtg tagtgccaat cagtgaagaa caaaattctg tgccaacatt
     9181 acaaaaaggt catcagaatg ctttgccaaa cccagagaag gaatttctat gtgaaaatgt
     9241 tttttcagag atgagccaaa attcttctct atcagaatcc ttgcccataa ctcagaaaat
     9301 atctgttggt cgactttccc ctcaacaaga atcttcagct tcgagtaaaa ggatgctttc
     9361 cccagcaaat agtttagaca ttgccatgga aaagcaccag aagcgggcca aagatgaaaa
     9421 tggagctgtt tgtgcaacag acgtgagacc tttagaggct ttgagttcga gagttaatga
     9481 agctagtaaa cagaagaagc ctattttagt gagacaggtt tgtactacag agcccctgga
     9541 cggtgtgatg ttggaaaagg atgttttttc tcaacctgaa attagtaatg aggctgttaa
     9601 tttgacaaat gttttaccag ctgataattc atcaacagga tgctctaaat ttgtcgttat
     9661 agaacctata agtgaattgc aggaatttga aaacatcaag tcatccacat cattaactct
     9721 tacagttcga agttcacctg ctccttcaga aaatactcat atttctcctt tgaaatgtac
     9781 agacaataac caagaaagga agtctccagg ggttaaaaat caaggtgaca aagtgaacat
     9841 ccaagagcaa agtcaacagc cagtcacttc tctttcattg tttaacatca aggacaccca
     9901 gcagctggct ttccctagcc tgaaaactac aaccaacttt acatggtgtt atctcttaag
     9961 gcagaagtcg ttgcatttgc ctcagaagga ccagaaaact tcagcctata ctgattggac
    10021 agtaagcgcc agtaatccaa atccactcgg tttgcccaca aaagttgcac ttgctctcct
    10081 taattcaaaa cagaacactg gaaaatcact atactgtcaa gcaataacta cccattccaa
    10141 gtcagactta ttggtctatt caagcaagtg gaaaagcagc ttaagcaaga gagcattagg
    10201 taatcaaaag tccacagtag ttgaattcag caataaagat gcctctgaaa ttaacagtga
    10261 gcaagataaa gaaaattcct taatcaaaag tgaaccaaga agaattaaaa tatttgatgg
    10321 aggatataag tcaaatgaag agtatgtata tgtccgaggc aggggaagag gaaaatacat
    10381 ttgtgaagaa tgtggaatac gttgtaagaa acctagcatg ttaaagaaac acatacgaac
    10441 ccatacagat gtccgcccct accactgcac ttactgtaac ttctccttta agactaaagg
    10501 aaatctgaca aaacacatga agtccaaggc acatagcaag aaatgtgtgg atttaggcgt
    10561 ctcagtaggt ttaatagatg aacaggatac agaagaatca gatgaaaaac agagattcag
    10621 ttatgagcga tctggatatg atcttgaaga atctgatggc ccagatgagg atgacaatga
    10681 aaatgaagac gatgatgagg acagccaggc tgaatcagtc ctgtcagcca caccctcagt
    10741 cacagctagc ccgcagcacc ttccatctag aagtagcctt caggaccctg tgagtactga
    10801 cgaggatgtc aggatcaccg attgcttttc tggggtacac acggacccaa tggacgttct
    10861 gcccagggcg ctgctcacca gaatgactgt cctgagcaca gcacagtctg actacaatag
    10921 gaagacactc tctccgggga aggccaggca gcgtgctgcg agagatgaaa acgacacaat
    10981 tccgtctgta gacacttcca ggtccccgtg tcatcagatg tctgtggact accctgagtc
    11041 agaagaaatt ctgagaagtt ctatggcagg aaaagctgtt gctataacac agagcccatc
    11101 atctgtaaga cttcctcctg ctgcagctga gcacagcccc cagacagcag cggggatgcc
    11161 ttctgtggcc tcaccacatc ctgaccctca agaacagaag cagcaaataa ctctacagcc
    11221 gactccaggc ttgccttctc cccacactca tttgtttagc caccttcctt tgcattccca
    11281 gcagcaatcg aggacacctt ataatatggt tccagttggg gggatccatg tggtacctgc
    11341 tggcctcaca tactccacgt ttgtgcccct tcaggctgga ccagtgcagc tcacgatccc
    11401 tgctgtcagt gtcgttcaca gaactttggg tactcatagg aatacggtca cagaagtgtc
    11461 tggcactaca aaccctgctg gagtggctga attaagcagt gttgtgccat gtattcctat
    11521 cggccaaatc cgcgtgccag gccttcagaa cctaagtacc ccaggcttgc agtcactccc
    11581 ctcgttaagc atggaaaccg tcaatattgt aggcctagcc aatacaaata tggccccaca
    11641 agtccatcca ccaggactgg ctctgaatgc tgtcggactg caggttctga ctgcaaaccc
    11701 ttcatcacaa agcagccccg cccctcaggc acacattcca ggtctccaga tcttgaacat
    11761 agcattgccc accttaatcc cctcagtcag tcaagtagcc gttgatgcac agggagctcc
    11821 agaaatgcca gcttcccaaa gcaaagcatg cgagacacaa cccaagcaga cttctgtagc
    11881 cagcgcaaac caggtcagca ggaccgagtc tcctcagggg ttacctacag tccagcggga
    11941 aaatgcaaaa aaagttctga atccacctgc ccctgcaggt gaccatgcaa ggcttgatgg
    12001 cctgagtaaa atggacacag agaaggctgc ctcggcaaat cacgtgaagc ccaagcctga
    12061 actcacttcc atacagggcc aaccagcgtc cacgtcacaa cctctgctga aggcacattc
    12121 tgaagttttt acaaagccct caggccagca gactctctct ccagacagac aggttcccag
    12181 gcccacagca ctaccgcgga ggcagcccac tgtgcacttc agcgacgtga gcagcgatga
    12241 tgacgaggac aggcttgtga tagcaaccta gtaatgaact agtccgtaac ttgaaagtat
    12301 ttcgatttct tggctttata tatcttgtgg aaaggacgat ctgcatacat tacggtgttg
    12361 tacacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
    12421 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
    12481 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
    12541 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
    12601 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
    12661 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
    12721 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
    12781 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
    12841 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
    12901 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
    12961 tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
    13021 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
    13081 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
    13141 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
    13201 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
    13261 aaatctctag cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg
    13321 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
    13381 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    13441 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
    13501 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
    13561 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
    13621 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
    13681 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
    13741 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
    13801 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
    13861 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
    13921 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
    13981 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
    14041 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
    14101 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
    14161 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
    14221 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
    14281 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
    14341 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
    14401 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
    14461 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
    14521 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
    14581 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
    14641 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
    14701 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
    14761 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
    14821 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
    14881 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
    14941 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
    15001 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
    15061 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
    15121 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
    15181 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
    15241 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
    15301 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
    15361 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
    15421 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
    15481 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
    15541 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
    15601 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
    15661 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
    15721 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
    15781 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
    15841 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
    15901 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
    15961 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
    16021 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
    16081 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
    16141 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
    16201 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
    16261 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
    16321 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
    16381 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
    16441 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
    16501 aacaaaagct ggagctgcaa gc
//
