LOCUS       sequence-353225-       16240 bp    DNA     circular SYN 15-DEC-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 16240)
  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 16240)
  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"; 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: TFORF2733
FEATURES             Location/Qualifiers
     source          1..16240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    9178..9766
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9231..9251)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9649..9660)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9838..10071
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10149..10270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10186..10205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10240..10259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10310..10445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10465..10484)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10466..10484)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10494..10511)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10494..10510)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10503..10525)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10652..11107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10739..10758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10949..10970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11133..11237
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11238..11306,11307..12098)
                     /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(11456..11475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12269..12857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12758..12777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13011..13028
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13145..13166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13181..13186,13187..13204,13205..13211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13219..13235
                     /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     13224..13246
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13243..13259
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13243..13259
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13278..13298
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13280..13298
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        13326..13552
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             13553..13733
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    13780..13905
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    14398..14631
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             14816..14860
                     /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        15091..15301
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        15113..15309
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      15160..15295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             15341..15943
                     /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(15341..15360)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     15837..15857
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    15986..16103
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gctagcaccg ccaccatgtt
     1201 ccgcagggca cgccttagcg tgaagccgaa tgtcaggcct ggtgtaggcg ccaggggctc
     1261 cacagcttcc aatccccagc gtggacggga gtctcccagg ccgccggatc ctgccacgga
     1321 ctctgcttcc aagcccgcgg agcccacaga tgtgcccaca gtcgatttcg gtggagcgga
     1381 gccccaagaa aaggctccta ggagcagtac tgaaaagact ggtggtgaca atgatgttga
     1441 agaatccagt agatcttcct ctactgtttc acagagaaga aagcgaatat caagtacttc
     1501 tagcctggtt aagtctagtg tcagtgttcc ttcagaatct catcccttat ctacaattaa
     1561 tcaagaggct ccacagccaa ctgccacttc aacaaaagag aaacagccat gctcagacag
     1621 ataccgaata tacaaagccc agaaactgag ggaaatgtta aaagaagaat tgagaaaaga
     1681 gaagaaacaa tggaaaaaca aatatgctat aaatgaaagt cagaggccac cagatcgttc
     1741 aaaaatgact atgagagact tcatatatta tctaccagat aataatccaa tgacttcttc
     1801 actggaacaa gaaaagaaaa ctgaaaagcc atcgactcca gtccagacaa gagagcaaga
     1861 aggtaagagt actcctaatg ctgaagataa tgaaatggaa gaagagacag atgatgggcc
     1921 attactggtt cctcgagtaa aagtggcaga agatggttcc attattttgg atgaagaaag
     1981 tttaactgta gaagttttaa gaacaaaagg cccttgtgtt gttgaagaaa atgaccccat
     2041 atttgagcgc ggttctacaa ctacatactc cagctttagg aaaaactatt actctaaacc
     2101 atggtcaaat aaagaaacag atatgttttt tttagccatc agcatggtag gaactgactt
     2161 ttctatgatc ggacaacttt ttcctcacag agcaaggata gaaattaaga ataaatttaa
     2221 acgggaagag aaaacaaatg gatggagaat agacaaagca ttccaggaaa agcgcccttt
     2281 tgacttcgat ttttttgctc atttgcttca gaaagttctt gctgaagaag agaaaagaaa
     2341 acaaaaatct gttaaaaatc acagtttaaa ggagaagaaa tccaccaaac cacggaaaaa
     2401 tgtaaaagtg aaaaaagttg cctgtgaagg agtgaataat gatccagatg agtctatgag
     2461 ttctagaatt tcagacacgg aaagatctca gaaggatgct cagacagttg aagaagagtc
     2521 tctgacctta tcaagggagg atgcagagca ggttgcatta gaagtagacc taaatcaaaa
     2581 gaaaagaagg aggaagaagc aagatggagc taatgaactg ggagtaaaca atcttttaga
     2641 aaatgccact gttcaggcgg gtccttctaa aggagaaaaa cacaagaata aatgtcaggc
     2701 tataaggcct gagctaaagg aaggtgaatg cagtaaggag cagatgcttt cctgcacaca
     2761 aaacatagat ggcattgtgg gttttgcctc cactgaaaaa gttgagaaaa gaactgaccc
     2821 catcctttca ttaagtaatc aacaagatgc cacatcagta gcaactgagt cttcagaatc
     2881 aagcacttca gatttgcctt cattcgaagt tggaattaga gcattgtgtg aggtgaataa
     2941 tgctgagggt agttgtatag aagaaagaaa tgttgaccta aaaaataatt cactggaaat
     3001 tgatcaaaca gaaaatgtta aaccaatgtt gagaggtcgc ttccaaagac ctaaacccaa
     3061 tttgtcaagg gctgggaaga aatcagttct ttcacaaggc aaaacagagt cagagagcaa
     3121 gaattcacat tcaaaaactt cagttgaaaa gaaccacgtg gaaaaagata aaatgaatac
     3181 attggacatt ttgagaatgg agactacaga gagagagaat ccagaagctg aaactgtatc
     3241 tgttttgggt gaaaaaaatt gtctgcagga agggagtcaa ctaaaggctt taagacctgt
     3301 acaagtgagg ggccgattgc aaaagccaaa gccaaatgca ggtaaagctg ctgaaagaaa
     3361 agaaattctc atatcacagg aagaaattgg ggccaatgta gagaagaatg aaaatgaatc
     3421 ctgtgctgat agagatactc ctcaacacat ggaagatcaa tcgcgtaaag attttgaaga
     3481 ggaagatgtc atattacagc ctgagaaaaa tgattctttt caaaatgtgc agccagatga
     3541 gcccaaggtt cttaatgaat gtctaagcgt tcaagagaat aataaggcaa ataaacttaa
     3601 ccaagtccca attctaagga ctcgatttca gaaaccaaag ccaaatatag gaagaggaac
     3661 tggaaggaga gaaatttcct caaaggaaga ggtactagag aagattcttg tctctgggga
     3721 aatggcggca gcattgagag aaactgtaag actagacacc tcaccaaagg agatggtacc
     3781 agcagagatt aatactaaag aaatgcagtc agatttaaaa gaaactggaa gaagagccat
     3841 ttctcccagg gagaagattc tagatgtgat tgatgacacc atagaaatgg agacaggtct
     3901 gaaagcaatg ggaagagaga tttgtctaag ggagaagacg ccagaggtga ttgatgccac
     3961 tgaggaaata gacaaagatt tggaagaagc tggaagaaga gaaatatccc cacagaaaaa
     4021 tggcccagag gaggttaagc ctctaggtga agtggagaca gatttgaaag caactggaaa
     4081 tgagagttcc ccaagggaga agacaccaga ggtgactgat gccactgagg aaatagacaa
     4141 aaatttggaa gaaactggaa gaagaaaaat atccccaagg gaaaatggcc cagaggaggt
     4201 caagcctgta gatgaaatgg agacagattt gaacgcaact ggaagagaga gttctccaag
     4261 ggagaagaca ccagaggtga ttgatgctac tgaggaaata gatttggaag aaactgaaag
     4321 agaagtatcc ccacaggaaa atggactaga ggaggtcaag cctctaggtg aaatggagac
     4381 ggatttgaaa gcaactggaa gagacagttt cccaaggggg aagacaccag aggtgattga
     4441 tgccattgag gaaatagaga tagatttgga agaaactgaa agagaaatat ccccacagga
     4501 aaatggccta gaggaggtta agcctctagg tgaaatgcaa acagatttga aagcaaccgg
     4561 aagggagatt tccccaaggg agaagacacc agaggtgatt gatgccactg aggaaataga
     4621 caaagatctg gaagaaactg gaagaagaga aatatcccca gaggaaaatg gcccagagga
     4681 ggtcaagcct gtagatgaaa tggagacaga cttgaaaaca actggaagag agggttcctc
     4741 aagggagaag acacgagagg tgattgatgc tgctgaggta atagagacag atttggaaga
     4801 aactgaaaga gaaatatcgc cacaggaaaa tggcccagag gaggtcaagc ctgtaggtaa
     4861 aatggagaca gatttgaaag aaattagaga agaaatttcc caaagggaaa aggtgctagc
     4921 agagttcagt gctataaggg aaaaggagat tgatttgaaa gaaactggaa aaagagacat
     4981 tcccatcatg gagaaagtat caggaaagat ggctgttgtt gaagaaatgg aggcagattt
     5041 gaaagaaact ggaaaagaaa attttagaga gagaggatct gaagagatct gtgttactga
     5101 ggaaaaggtg gcagaattga aacaaactgg aaaaacagac atttctccaa gggaaaacga
     5161 gctagaggag accagtacct caagacaaac tgacacacat ttaatgcaga gcggtagcaa
     5221 tgacttcagt gctgtgcctt cactagatat tcagaacatt agcagtgaag tactgtcgat
     5281 gatgcataca cctgtagaag aaaaaagaaa ttctgaaaaa gaagtatcaa gtcacttcag
     5341 tcatttcaag atttcttcac agactcatga atctgataaa acagaagtcc aggggattca
     5401 atctccagat gttccagagc agttttcaga tattaattta agcaaatctc ttcctcaaga
     5461 acagaagcca cttgaaatta aaccagcacc ttttgtgagg agccgattca aaagaccaaa
     5521 accaaactta gcaagagcag ctttgaagag agagactaca gaatcagaaa aatatatata
     5581 tgagaagaaa tcagaaacca agaaaatgga gactattgtg atgcaagaaa ataatgaaca
     5641 aactgatact ctcccttctc aacatgatga agcttcccta atgatatcaa gagaaaaaga
     5701 cacattaggt cacaggaatg aggaggctgt gatattgcca tgtacacaga ctgaaaggaa
     5761 cctttcacct tcaaattctt gtgaacctaa agaggagtct cagtcagcac cagtccagaa
     5821 aaatgactca gttgtttctg tggggactaa taatgtaaac actttccagc aagaaatgaa
     5881 ggaaagtgtt atccaaactg ctcgacaagt aaggggccga cttcagagac cgagaccaaa
     5941 tataagaaag acaggacaga ggcaaatagt agacaaaggt gaagccaaag gcataattaa
     6001 ggaaggaaga acgatattac caaaagatga aactgaaaag aaagtcttaa ctgtgtcaaa
     6061 ttctcaaatt gaaactgaaa ttgaagttcc atcgtccgca gttccagaac acagaatgta
     6121 tgaaaatcaa agtcaggtgg ttcttgtaga aaaccttcat gttaacaaaa caaatgaaac
     6181 aatcagacat gaaaataaac cgtatgttcc tagttcagca caaatgacaa gaaggaaatt
     6241 ccaaaaggct aagccaaatt tgggaagagc acacagtaag aaagaggaac cagttttaga
     6301 aaaagtcaca acagatcaga gcaaggaagg caagccagaa gatcatttgc tgcagaaagg
     6361 agcttccaac acccagctcc ttctaaaaga aaaagctgag cttctgacat ctctggaggt
     6421 ttcagcaaga aaagattgtg taggttccaa agagtctgct ttggcaaaaa tagatgcgga
     6481 attagaagaa gttggaccat caagaagggt tggagaggaa actgtaggag ataattcacc
     6541 atcttcagtt gttgaagagc aatatctcaa taaactaaca agctgtccac aaccgttaaa
     6601 cgaaacaagt tactctaaaa ttgccctgga tgggaaaaca actatctctt ctacatctga
     6661 gtatgagaga aatcgtggtg aaaggagaag tcataaaaag ttcaaaccaa atgtcaccag
     6721 aggtcgtgga tcaaaacgag ttcggggtaa gacctctaag aaggaaccta gagcttccaa
     6781 ggccatgctg gtgactcttc gggcttccca ggaagaagat gatgatgctg acgattttga
     6841 gtctgactat gaggaagaaa gctatcatct tgctcccgaa gaagtaaaca aagctccagt
     6901 atttgtacct gttggtctca gatctcctga acctgtttct gctcagattg aggaaacaat
     6961 ggaagagctt gaaataactg tgaatgtccc agatgtagga tgcatagctg ttgttgaaca
     7021 tgagctacca aacacagatg tgactactga agaaatgaaa caagaagaaa atttgagtgt
     7081 accgtttgaa atgaccacaa gtgaacatat ccaagatgaa ccaggtacca atgatggaag
     7141 caccgaagct gcaataactt tacttacaat gggagatcta gtattgcagt cagagatcag
     7201 tagtgaacag ggtgatgtag gagtatgtat aattcctcat gttcattcaa aggataaaag
     7261 ccatattcct tctagcctag ataatgtaaa tcacaaaatt gttcatgaat gtcaggaact
     7321 ttcttcacct gtcattacta catctcctgc atcatttgaa gaaaacaaga ttgtattgga
     7381 ggaacaaagt tccagagaag aaataagttt aatggagaaa gtaaaggaga atgctacacc
     7441 aaccaggaat acaatttcta aagtgaccag taatttgaga ataagaagta ggcttgctaa
     7501 gcctaaacca aatcttgaga agactttagg gaccaacagg cttgatgatt atcaggaagt
     7561 ttccagtttg tgtgtaacca aaggggcaga aatggaaact caaagagaaa cagagaaaaa
     7621 tgcttccaaa gcaacagaat tggaaaataa aaacctcgga ccagttacaa cagcagagaa
     7681 taaggatcag agcaaattgg catgtgtaca tggtatcaaa gggaccagta tttcttcaga
     7741 agtaaaccta actgaaagaa acgaaaatca agaagagagc tctcaggagg ttcacatgtt
     7801 gtcagttgct ccagttgctt cctctgagac agggccctgc acacttggtt tggatagggg
     7861 tcttggtgaa aattctgttg aagagcccca gataaaggac tctaaaggag acagtgtgct
     7921 tacacttcct gtgccagagt atacaccaac aagtattcca gaagtccaac aagagaatat
     7981 aatcaatcct caagacctaa cagtgaatct agttgctaat gtacctcaag atggagaaga
     8041 tgaacaagcc tttattttaa ctctggtgga aatcccagcc aatgcagtag aagaatttac
     8101 tgatgccact gcacagttca tgccaaaccc tttactgcca gctcccatat tggtcaaatc
     8161 agtgaatacc gaagaaaggg gtgacatgag tatttgttta ccagcaactt cagttggtca
     8221 agatgccatg ggtttatcta tttctggaag agataattct aaaaagccgc ctgataattt
     8281 ggatcttgta tctaggaaga gatttcaatg caggcttgat aaaaatgacc acattcctcc
     8341 tgccaaaaaa cgttcactca ctttaagaga tgactgtcaa gaatatacca ctgaggtgca
     8401 ctcaaaggaa ttaacaaatg tttttgagga aacaggggag tctcacaagg gacaagatat
     8461 ttttcttacc tcaggaagca cactgacaac tccagaacct caaagacagc aagttgaagc
     8521 agcttttcag agtagaggat ctagatctcc tgatgcatgc atggacaaga atgtgcctca
     8581 gttacctcag gatgaaatga ttgtgtctga taaggaagaa agaactgatg ctgctcctaa
     8641 gtctcagcaa atggatagca gaacatcgtc ttctaaagcc tcactatcca gacctggcag
     8701 aagacccctg ggatttttat ctttaatatg ctcaaagaat agtttggagt ctgatgaacc
     8761 tatgcaagtc catagtaaga aacgcctaaa acctcttata cctggattaa gaaagaaatt
     8821 gaaaagatct aatccattca atgaaagcca ggaaaaaaat cgagagtcct ctgatctgct
     8881 tccatctcca agtgttatta ctactcaatc tgagaatatt agcagctcag caactcaggt
     8941 ttcttgtgat cagcccttac tgaaagaagg atataaaagt gcccaaaagc gggcccctca
     9001 aggggaggca accacagtct ctgaatattt cttcaatgat atcttcattg aagtggatga
     9061 aacagaatag taatgaacta gtccgtaact tgaaagtatt tcgatttctt ggctttatat
     9121 atcttgtgga aaggacgaga taccgaggcc aaacagcaat atacgcgtta agtcgacaat
     9181 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     9241 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     9301 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     9361 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     9421 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     9481 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     9541 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     9601 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     9661 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     9721 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     9781 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     9841 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     9901 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     9961 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
    10021 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
    10081 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
    10141 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
    10201 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
    10261 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
    10321 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
    10381 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
    10441 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
    10501 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
    10561 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
    10621 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
    10681 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
    10741 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
    10801 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
    10861 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
    10921 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
    10981 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
    11041 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
    11101 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
    11161 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
    11221 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
    11281 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
    11341 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
    11401 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
    11461 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
    11521 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
    11581 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
    11641 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
    11701 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
    11761 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
    11821 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
    11881 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
    11941 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
    12001 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
    12061 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
    12121 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
    12181 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
    12241 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
    12301 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
    12361 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
    12421 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    12481 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
    12541 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
    12601 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
    12661 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
    12721 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
    12781 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
    12841 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
    12901 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
    12961 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
    13021 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
    13081 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
    13141 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
    13201 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
    13261 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
    13321 cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
    13381 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
    13441 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
    13501 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
    13561 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
    13621 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
    13681 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
    13741 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
    13801 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
    13861 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
    13921 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
    13981 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
    14041 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
    14101 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
    14161 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
    14221 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
    14281 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
    14341 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
    14401 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
    14461 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
    14521 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
    14581 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
    14641 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
    14701 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
    14761 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
    14821 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
    14881 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
    14941 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
    15001 gtttcagacc cacctcccaa ccccgagggg acccatgcat tgcatctcaa ttagtcagca
    15061 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcgtctc
    15121 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
    15181 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    15241 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    15301 ttttgcaaaa agctttctag aggtaccacc ggagcttacc atgaccgagt acaagcccac
    15361 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
    15421 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
    15481 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
    15541 ggacgacggc gccgccgtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
    15601 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
    15661 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
    15721 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
    15781 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
    15841 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
    15901 accgcgcacc tggtgcatga cccgcaagcc cggtgccagt taacgaattc tagatcttga
    15961 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
    16021 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
    16081 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc
    16141 gccggctcga ggcctgcagg tgcaaagatg gataaagttt taaacagaga ggaatctttg
    16201 cagctaatgg accttctagg tcttgaaagg agtgggaatt
//
