LOCUS       sequence-396219-       14248 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 14248)
  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 14248)
  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: TFORF2847
FEATURES             Location/Qualifiers
     source          1..14248
                     /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"
     misc_feature    10105..10693
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10158..10178)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10576..10587)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10765..10998
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    11076..11197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11113..11132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11167..11186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      11237..11372
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(11392..11411)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11393..11411)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11421..11438)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11421..11437)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11430..11452)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11579..12034
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11666..11685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11876..11897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        12060..12164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12165..12233,12234..13025)
                     /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(12383..12402)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13196..13784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13685..13704
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13938..13955
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    14072..14093
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(14108..14113,14114..14131,14132..14138)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14146..14162
                     /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     14151..14173
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     14170..14186
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14170..14186
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     14205..14225
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        14207..14225
                     /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 caccatgcag tcctttcggg agcaaagcag
     4141 ttaccacgga aaccagcaaa gctacccaca ggaggtacac ggctcatccc ggctagaaga
     4201 gttcagccct cgtcaggccc agatgttcca gaattttgga ggtacaggtg gcagtagtgg
     4261 cagcagtggc agtggcagtg gtggtggacg acgaggagca gcagctgctg cggcagcgat
     4321 ggctagcgag acctctggcc atcaaggtta ccagggtttc aggaaagagg ctggagattt
     4381 ttactacatg gcaggcaaca aagaccccgt gactacagga accccacagc ctcctcagcg
     4441 aaggccttct gggcctgtgc agagctatgg acccccccag gggagcagct ttggcaatca
     4501 gtatgggagt gagggtcatg tgggccagtt tcaagcacag cactctggcc ttggcggtgt
     4561 gtcacattat cagcaggatt acactgggcc tttctctcca gggagtgctc agtaccaaca
     4621 gcaggcttcc agccagcagc agcagcagca agtccagcag ttgagacaac agctttacca
     4681 gtcccatcag cccctgccac aggccactgg ccaaccagca tccagctcat cccatctaca
     4741 gccaatgcag cggccctcaa ctctgccatc ctctgctgct ggttaccagt taagagtggg
     4801 tcagtttggc caacactatc agtcttctgc ttcctcctcc tcctcctcct ccttcccttc
     4861 accacagcgt tttagccagt ctggacagag ctatgatggc agttacaatg tgaatgctgg
     4921 atctcagtat gaaggacaca atgtgggttc taatgcacag gcttatggaa cacaatccaa
     4981 ttacagctat cagcctcaat ctatgaagaa ttttgaacag gcaaagattc cacaagggac
     5041 ccaacagggg cagcagcagc agcaaccgca gcaacaacaa cacccttctc agcatgtgat
     5101 gcagtatact aacgctgcca ccaagctgcc cctgcaaagc caagtggggc agtacaacca
     5161 gcctgaggtt cctgtgaggt cccccatgca gtttcaccag aacttcagcc ccatttctaa
     5221 cccttctcca gctgcctctg tggttcagtc tccaagctgt agttctaccc catctcctct
     5281 catgcagact ggggagaatc tccagtgtgg gcaaggcagt gtgcctatgg gttccagaaa
     5341 cagaatttta cagttaatgc ctcaactcag tccaacccca tcaatgatgc ccagtcctaa
     5401 ttctcatgct gcaggcttca aagggtttgg actagaaggg gtaccagaaa agcgactgac
     5461 agatcctggg ttgagtagtt tgagtgctct gagtactcaa gtggccaatc ttcctaacac
     5521 tgtccagcac atgttacttt ctgatgccct gactcctcag aagaagacct ccaagaggcc
     5581 ctcatcttcc aagaaagcag atagctgcac aaattctgaa ggctcctcac aacctgaaga
     5641 acagctgaag tcccctatgg cagagtcatt agatggaggc tgctccagca gttcagagga
     5701 tcaaggcgag agagtgcggc aactaagtgg ccagagcacc agctctgaca ccacctacaa
     5761 gggtggagcc tctgagaaag ctggctcctc accggcacaa ggtgctcaga atgaaccccc
     5821 cagactcaat gctagtcctg ccgcaagaga agaggccacc tcaccaggcg ctaaggacat
     5881 gccattgtca tccgacggga acccaaaggt taatgagaag actgttgggg tgattgtctc
     5941 ccgggaagcc atgacaggtc gggtagaaaa gcctggtgga caagataaag gctcccaaga
     6001 ggatgatcct gcagccactc aaaggccacc tagcaatggt ggggcaaagg aaaccagtca
     6061 tgcatcactt ccccagccag agcctccagg aggaggaggg agcaaaggaa acaagaatgg
     6121 cgataacaac tccaaccata atggagaagg aaatggccag agtggccact ctgcagcggg
     6181 ccctggtttt acgagcagaa ctgagcctag caaatctcct ggaagtctgc gctatagtta
     6241 caaagatagt ttcgggtcag ccgtgccacg aaatgtcagt ggctttcctc agtatcctac
     6301 agggcaagaa aagggagatt tcactggcca tggggaacga aagggtagaa atgaaaaatt
     6361 cccaagcctc ctgcaggaag tgcttcaggg ttaccaccac caccctgaca ggagatattc
     6421 taggagtact caagagcatc aggggatggc tggtagccta gaaggaacca caaggcccaa
     6481 tgtcttggtt agtcaaacca atgaattagc tagcaggggc cttctgaaca aaagcattgg
     6541 gtctctatta gaaaatcccc actggggccc ctgggaaagg aaatcaagca gcacagctcc
     6601 tgaaatgaaa cagatcaatt tgactgacta tccaattccc agaaagtttg aaatagagcc
     6661 tcagtcatca gcacatgagc ctgggggttc cctctctgaa agaagatcag tgatctgtga
     6721 tatttctcca ctaagacaga ttgtcaggga cccaggggct cactcactgg gacacatgag
     6781 tgccgacacc agaattggga ggaatgaccg tctcaatcca actttaagtc agtcggtcat
     6841 tcttcctggt ggtttggtgt ccatggaaac caagctgaaa tcccagagcg ggcagataaa
     6901 agaggaagac tttgaacagt ctaaatctca agctagtttc aacaacaaga aatctggaga
     6961 ccactgccat cctcctagca tcaagcatga gtcttaccgc ggcaatgcca gccctggagc
     7021 agcaacccat gattcccttt cagactatgg cccgcaagac agcagaccca cgccaatgcg
     7081 gcgggtccct ggcagagttg gtggtcggga gggcatgagg ggtcggtccc cttctcaata
     7141 tcatgacttt gcagaaaaat tgaaaatgtc tcctgggcgg agcagaggcc cagggggaga
     7201 ccctcatcac atgaatccac acatgacctt ttcagagagg gctaaccgga gttctttaca
     7261 cactcccttt tctcccaact cagaaaccct ggcctctgct tatcatgcaa atactcgggc
     7321 tcatgcttat ggggacccta acgcaggttt gaattctcag ctgcattata agagacagat
     7381 gtaccaacag caaccagagg agtataaaga ctggagcagc ggttctgctc agggagtaat
     7441 tgctgcagca cagcacaggc aggaggggcc acggaagagt ccaaggcagc agcagtttct
     7501 tgacagagta cggagccctc tgaaaaatga caaagatggt atgatgtatg gcccaccagt
     7561 ggggacttac catgacccca gtgcccagga ggctgggcgc tgcctaatgt ctagtgatgg
     7621 tctgcctaac aagggcatgg aattaaagca tggctcccag aagttacaag aatcctgttg
     7681 ggatctttct cggcaaactt ctccagccaa aagcagcggt cctccaggaa tgtccagtca
     7741 aaaaaggtat gggccgcccc atgagactga tggacatgga ctagctgagg ctacacagtc
     7801 atccaaacct ggtagtgtta tgctgagact tccaggccag gaggatcatt cttctcaaaa
     7861 ccccttaatc atgaggaggc gtgttcgttc ttttatctct cccattccca gtaagagaca
     7921 gtcacaagat gtaaagaaca gtagcactga agataaaggt cgcctccttc actcatcaaa
     7981 agaaggcgct gataaagcat tcaattccta tgcccatctt tctcacagtc aggatatcaa
     8041 gtctatccct aagagagatt cctccaagga ccttccaagt ccagatagta gaaactgccc
     8101 tgctgttacc ctcacaagcc ctgctaagac caaaatactg cccccacgga aaggacgggg
     8161 attgaaattg gaagctatag ttcagaagat tacatcccca aatattagga ggagcgcatc
     8221 ttcgaacagt gcggaggctg ggggagacac ggttacgctt gatgatatac tgtctttgaa
     8281 gagtggtcct cctgaaggtg ggagtgttgc tgttcaggat gctgacatag agaagagaaa
     8341 aggtgaggtg gcttcggacc tagtcagtcc agcaaaccag gagttgcacg tagagaaacc
     8401 tcttccaagg tcttcagaag agtggcgtgg cagcgtggat gacaaagtga agacagagac
     8461 acatgcagaa acagttactg ccggaaagga accccctggt gccatgacat ccacaacctc
     8521 acagaagcct ggtagtaacc aagggagacc agatggttcc ctgggtggaa cagcaccttt
     8581 aatctttcca gactcaaaga atgtacctcc agtgggcata ttggcccctg aggcaaaccc
     8641 caaggctgaa gagaaggaga acgatacagt gacgatttca ccgaagcaag agggtttccc
     8701 tccaaaggga tatttcccat caggaaagaa gaaggggaga cccattggta gtgtgaataa
     8761 gcaaaagaaa cagcagcagc caccgcctcc accccctcag cccccacaga taccagaagg
     8821 ttctgcagat ggagagccaa agccaaaaaa acagaggcaa aggagggaga gaaggaagcc
     8881 tggggcccag ccgaggaagc gaaaaaccaa acaagcagtt cccattgtgg aaccccaaga
     8941 acctgagatc aaactaaaat atgccaccca gccactggat aaaactgatg ccaagaacaa
     9001 gtctttttac ccttacatcc atgtagtaaa taagtgtgaa cttggagccg tttgtacaat
     9061 catcaatgct gaggaagaag aacagaccaa attagtgagg ggcaggaagg gtcagaggtc
     9121 actgacccct ccacctagca gcactgaaag caaggcgctc ccggcctcgt cctttatgct
     9181 gcagggacct gttgtgacag agtcttcggt tatggggcac ctggtttgct gtctgtgtgg
     9241 caagtgggcc agttaccgga acatgggtga cctctttgga cctttttatc cccaagatta
     9301 tgcagccact ctcccgaaga atccacctcc taagagggcc acagaaatgc agagcaaagt
     9361 taaggtacgg cacaaaagtg cttctaatgg ctccaagacg gacactgagg aggaggaaga
     9421 gcagcagcag cagcagaagg agcagagaag cctggccgca caccccaggt ttaagcggcg
     9481 ccaccgctcg gaagactgtg gtggaggccc tcggtccctg tccagggggc tcccttgtaa
     9541 aaaagcagcc actgagggca gcagtgaaaa gactgttttg gactcgaagc cctccgtgcc
     9601 caccacttca gaaggtggcc ctgagctgga gttacaaatc cctgaactac ctcttgacag
     9661 caatgaattt tgggtccatg agggttgtat tctctgggcc aatggaatct acctggtttg
     9721 tggcaggctc tatggcctgc aggaagcgct ggaaatagcc agagagatga aatgttccca
     9781 ctgccaggag gcaggcgcca ccttgggctg ctacaacaaa ggctgctcct tccgatacca
     9841 ttacccgtgt gccattgatg cagattgttt gctacatgag gagaacttct cggtgaggtg
     9901 ccctaagcac aagcctcccc ttccgtgccc tctccccccc ttgcagaaca agaccgcgaa
     9961 aggcagcctc agcacagagc agtcggagcg ggggtagtaa tgaactagtc cgtaacttga
    10021 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaactta gacgaaagac
    10081 atatccatca cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
    10141 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
    10201 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
    10261 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
    10321 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
    10381 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
    10441 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
    10501 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
    10561 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
    10621 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
    10681 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
    10741 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
    10801 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
    10861 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
    10921 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
    10981 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
    11041 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
    11101 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
    11161 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
    11221 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
    11281 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
    11341 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
    11401 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
    11461 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
    11521 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
    11581 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    11641 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    11701 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    11761 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    11821 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    11881 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    11941 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    12001 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
    12061 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
    12121 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
    12181 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
    12241 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
    12301 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
    12361 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
    12421 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
    12481 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
    12541 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
    12601 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
    12661 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
    12721 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
    12781 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
    12841 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
    12901 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
    12961 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
    13021 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
    13081 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
    13141 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
    13201 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
    13261 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
    13321 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
    13381 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
    13441 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
    13501 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
    13561 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
    13621 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
    13681 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    13741 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
    13801 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
    13861 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
    13921 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
    13981 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
    14041 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
    14101 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
    14161 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
    14221 aaagggaaca aaagctggag ctgcaagc
//
