LOCUS       sequence-353411-       16885 bp    DNA     circular SYN 14-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 16885)
  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 16885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: TFORF1618
FEATURES             Location/Qualifiers
     source          1..16885
                     /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"
     regulatory      1190..1199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    9823..10411
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9876..9896)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10294..10305)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10483..10716
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10794..10915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10831..10850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10885..10904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10955..11090
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(11110..11129)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11111..11129)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11139..11156)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11139..11155)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11148..11170)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11297..11752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11384..11403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11594..11615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11778..11882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11883..11951,11952..12743)
                     /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(12101..12120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12914..13502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13403..13422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13656..13673
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13790..13811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13826..13831,13832..13849,13850..13856)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13864..13880
                     /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     13869..13891
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13888..13904
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13888..13904
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13923..13943
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13925..13943
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        13971..14197
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             14198..14378
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    14425..14550
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    15043..15276
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             15461..15505
                     /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        15736..15946
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        15758..15954
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      15805..15940
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             15986..16588
                     /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(15986..16005)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     16482..16502
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    16631..16748
                     /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 ccaccatggc
     1201 cgcgcacagg ccggtggaat gggtccaggc cgtggtcagc cgcttcgacg agcagcttcc
     1261 aataaaaaca ggacagcaga acacacatac caaagtcagt actgagcaca acaaggaatg
     1321 tctaatcaat atttccaaat acaagttttc tttggttata agcggcctca ctactatttt
     1381 aaagaatgtt aacaatatga gaatatttgg agaagctgct gaaaaaaatt tatatctctc
     1441 tcagttgatt atattggata cactggaaaa atgtcttgct gggcaaccaa aggacacaat
     1501 gagattagat gaaacgatgc tggtcaaaca gttgctgcca gaaatctgcc attttcttca
     1561 cacctgtcgt gaaggaaacc agcatgcagc tgaacttcgg aattctgcct ctggggtttt
     1621 attttctctc agctgcaaca acttcaatgc agtctttagt cgcatttcta ccaggttaca
     1681 ggaattaact gtttgttcag aagacaatgt tgatgttcat gatatagaat tgttacagta
     1741 tatcaatgtg gattgtgcaa aattaaaacg actcctgaag gaaacagcat ttaaatttaa
     1801 agccctaaag aaggttgcgc agttagcagt tataaatagc ctggaaaagg cattttggaa
     1861 ctgggtagaa aattatccag atgaatttac aaaactgtac cagatcccac agactgatat
     1921 ggctgaatgt gcagaaaagc tatttgactt ggtggatggt tttgctgaaa gcaccaaacg
     1981 taaagcagca gtttggccac tacaaatcat tctccttatc ttgtgtccag aaataatcca
     2041 ggatatatcc aaagacgtgg ttgatgaaaa caacatgaat aagaagttat ttctggacag
     2101 tctacgaaaa gctcttgctg gccatggagg aagtaggcag ctgacagaaa gtgctgcaat
     2161 tgcctgtgtc aaactgtgta aagcaagtac ttacatcaat tgggaagata actctgtcat
     2221 tttcctactt gttcagtcca tggtggttga tcttaagaac ctgcttttta atccaagtaa
     2281 gccattctca agaggcagtc agcctgcaga tgtggatcta atgattgact gccttgtttc
     2341 ttgctttcgt ataagccctc acaacaacca acactttaag atctgcctgg ctcagaattc
     2401 accttctaca tttcactatg tgctggtaaa ttcactccat cgaatcatca ccaattccgc
     2461 attggattgg tggcctaaga ttgatgctgt gtattgtcac tcggttgaac ttcgaaatat
     2521 gtttggtgaa acacttcata aagcagtgca aggttgtgga gcacacccag caatacgaat
     2581 ggcaccgagt cttacattta aagaaaaagt aacaagcctt aaatttaaag aaaaacctac
     2641 agacctggag acaagaagct ataagtatct tctcttgtcc atggtgaaac taattcatgc
     2701 agatccaaag ctcttgcttt gtaatccaag aaaacagggg cccgaaaccc aaggcagtac
     2761 agcagaatta attacagggc tcgtccaact ggtccctcag tcacacatgc cagagattgc
     2821 tcaggaagca atggaggctc tgctggttct tcatcagtta gatagcattg atttgtggaa
     2881 tcctgatgct cctgtagaaa cattttggga gattagctca caaatgcttt tttacatctg
     2941 caagaaatta actagtcatc aaatgcttag tagcacagaa attctcaagt ggttgcggga
     3001 aatattgatc tgcaggaata aatttcttct taaaaataag caggcagata gaagttcctg
     3061 tcactttctc cttttttacg gggtaggatg tgatattcct tctagtggaa ataccagtca
     3121 aatgtccatg gatcatgaag aattactacg tactcctgga gcctctctcc ggaagggaaa
     3181 agggaactcc tctatggata gtgcagcagg atgcagcgga acccccccga tttgccgaca
     3241 agcccagacc aaactagaag tggccctgta catgtttctg tggaaccctg acactgaagc
     3301 tgttctggtt gccatgtcct gtttccgcca cctctgtgag gaagcagata tccggtgtgg
     3361 ggtggatgaa gtgtcagtgc ataacctctt gcccaactat aacacattca tggagtttgc
     3421 ctctgtcagc aatatgatgt caacaggaag agcagcactt cagaaaagag tgatggcact
     3481 gctgaggcgc attgagcatc ccactgcagg aaacactgag gcttgggaag atacacatgc
     3541 aaaatgggaa caagcaacaa agctaatcct taactatcca aaagccaaaa tggaagatgg
     3601 ccaggctgct gaaagccttc acaagaccat tgttaagagg cgaatgtccc atgtgagtgg
     3661 aggaggatcc atagatttgt ctgacacaga ctccctacag gaatggatca acatgactgg
     3721 cttcctttgt gcccttgggg gagtgtgcct ccagcagaga agcaattctg gcctggcaac
     3781 ctatagccca cccatgggtc cagtcagtga acgtaagggt tctatgattt cagtgatgtc
     3841 ttcagaggga aacgcagata cacctgtcag caaatttatg gatcggctgt tgtccttaat
     3901 ggtgtgtaac catgagaaag tgggacttca aatacggacc aatgttaagg atctggtggg
     3961 tctagaattg agtcctgctc tgtatccaat gctatttaac aaattgaaga ataccatcag
     4021 caagtttttt gactcccaag gacaggtttt attgactgat accaatactc aatttgtaga
     4081 acaaaccata gctataatga agaacttgct agataatcat actgaaggca gctctgaaca
     4141 tctagggcaa gctagcattg aaacaatgat gttaaatctg gtcaggtatg ttcgtgtgct
     4201 tgggaatatg gtccatgcaa ttcaaataaa aacgaaactg tgtcaattag ttgaagtaat
     4261 gatggcaagg agagatgacc tctcattttg ccaagagatg aaatttagga ataagatggt
     4321 agaatacctg acagactggg ttatgggaac atcaaaccaa gcagcagatg atgatgtaaa
     4381 atgtcttaca agagatttgg accaggcaag catggaagca gtagtttcac ttctagctgg
     4441 tctccctctg cagcctgaag aaggagatgg tgtggaattg atggaagcca aatcacagtt
     4501 atttcttaaa tacttcacat tatttatgaa ccttttgaat gactgcagtg aagttgaaga
     4561 tgaaagtgcg caaacaggtg gcaggaaacg tggcatgtct cggaggctgg catcactgag
     4621 gcactgtacg gtccttgcaa tgtcaaactt actcaatgcc aacgtagaca gtggtctcat
     4681 gcactccata ggcttaggtt accacaagga tctccagaca agagctacat ttatggaagt
     4741 tctgacaaaa atccttcaac aaggcacaga atttgacaca cttgcagaaa cagtattggc
     4801 tgatcggttt gagagattgg tggaactggt cacaatgatg ggtgatcaag gagaactccc
     4861 tatagcgatg gctctggcca atgtggttcc ttgttctcag tgggatgaac tagctcgagt
     4921 tctggttact ctgtttgatt ctcggcattt actctaccaa ctgctctgga acatgttttc
     4981 taaagaagta gaattggcag actccatgca gactctcttc cgaggcaaca gcttggccag
     5041 taaaataatg acattctgtt tcaaggtata tggtgctacc tatctacaaa aactcctgga
     5101 tcctttatta cgaattgtga tcacatcctc tgattggcaa catgttagct ttgaagtgga
     5161 tcctaccagg ttagaaccat cagagagcct tgaggaaaac cagcggaacc tccttcagat
     5221 gactgaaaag ttcttccatg ccatcatcag ttcctcctca gaattccccc ctcaacttcg
     5281 aagtgtgtgc cactgtttat accaggcaac ttgccactcc ctactgaata aagctacagt
     5341 aaaagaaaaa aaggaaaaca aaaaatcagt ggttagccag cgtttccctc agaacagcat
     5401 cggtgcagta ggaagtgcca tgttcctcag atttatcaat cctgccattg tctcaccgta
     5461 tgaagcaggg attttagata aaaagccacc acctagaatc gaaaggggct tgaagttaat
     5521 gtcaaagata cttcagagta ttgccaatca tgttctcttc acaaaagaag aacatatgcg
     5581 gcctttcaat gattttgtga aaagcaactt tgatgcagca cgcaggtttt tccttgatat
     5641 agcatctgat tgtcctacaa gtgatgcagt aaatcatagt ctttccttca taagtgacgg
     5701 caatgtgctt gctttacatc gtctactctg gaacaatcag gagaaaattg ggcagtatct
     5761 ttccagcaac agggatcata aagctgttgg aagacgacct tttgataaga tggcaacact
     5821 tcttgcatac ctgggtcctc cagagcacaa acctgtggca gatacacact ggtccagcct
     5881 taaccttacc agttcaaagt ttgaggaatt tatgactagg catcaggtac atgaaaaaga
     5941 agaattcaag gctttgaaaa cgttaagtat tttctaccaa gctgggactt ccaaagctgg
     6001 gaatcctatt ttttattatg ttgcacggag gttcaaaact ggtcaaatca atggtgattt
     6061 gctgatatac catgtcttac tgactttaaa gccatattat gcaaagccat atgaaattgt
     6121 agtggacctt acccataccg ggcctagcaa tcgctttaaa acagactttc tctctaagtg
     6181 gtttgttgtt tttcctggct ttgcttacga caacgtctcc gcagtctata tctataactg
     6241 taactcctgg gtcagggagt acaccaagta tcatgagcgg ctgctgactg gcctcaaagg
     6301 tagcaaaagg cttgttttca tagactgtcc tgggaaactg gctgagcaca tagagcatga
     6361 acaacagaaa ctacctgctg ccaccttggc tttagaagag gacctgaagg tattccacaa
     6421 tgctctcaag ctagctcaca aagacaccaa agtttctatt aaagttggtt ctactgctgt
     6481 ccaagtaact tcagcagagc gaacaaaagt cctagggcaa tcagtctttc taaatgacat
     6541 ttattatgct tcggaaattg aagaaatctg cctagtagat gagaaccagt tcaccttaac
     6601 cattgcaaac cagggcacgc cgctcacctt catgcaccag gagtgtgaag ccattgtcca
     6661 gtctatcatt catatccgga cccgctggga actgtcacag cccgactcta tcccccaaca
     6721 caccaagatt cggccaaaag atgtccctgg gacactgctc aatatcgcat tacttaattt
     6781 aggcagttct gacccgagtt tacggtcagc tgcctataat cttctgtgtg ccttaacttg
     6841 tacctttaat ttaaaaatcg agggccagtt actagagaca tcaggtttat gtatccctgc
     6901 caacaacacc ctctttattg tctctattag taagacactg gcagccaatg agccacacct
     6961 cacgttagaa tttttggaag agtgtatttc tggatttagc aaatctagta ttgaattgaa
     7021 acacctttgt ttggaataca tgactccatg gctgtcaaat ctagttcgtt tttgcaagca
     7081 taatgatgat gccaaacgac aaagagttac tgctattctt gacaagctga taacaatgac
     7141 catcaatgaa aaacagatgt acccatctat tcaagcaaaa atatggggaa gccttgggca
     7201 gattacagat ctgcttgatg ttgtactaga cagtttcatc aaaaccagtg caacaggtgg
     7261 cttgggatca ataaaagctg aggtgatggc agatactgct gtagctttgg cttctggaaa
     7321 tgtgaaattg gtttcaagca aggttattgg aaggatgtgc aaaataattg acaagacatg
     7381 cttatctcca actcctactt tagaacaaca tcttatgtgg gatgatattg ctattttagc
     7441 acgctacatg ctgatgctgt ccttcaacaa ttcccttgat gtggcagctc atcttcccta
     7501 cctcttccac gttgttactt tcttagtagc cacaggtccg ctctccctta gagcttccac
     7561 acatggactg gtcattaata tcattcactc tctgtgtact tgttcacagc ttcattttag
     7621 tgaagagacc aagcaagttt tgagactcag tctgacagag ttctcattac ccaaatttta
     7681 cttgctgttt ggcattagca aagtcaagtc agctgctgtc attgccttcc gttccagtta
     7741 ccgggacagg tcattctctc ctggctccta tgagagagag acttttgctt tgacatcctt
     7801 ggaaacagtc acagaagctt tgttggagat catggaggca tgcatgagag atattccaac
     7861 gtgcaagtgg ctggaccagt ggacagaact agctcaaaga tttgcattcc aatataatcc
     7921 atccctgcaa ccaagagctc ttgttgtctt tgggtgtatt agcaaacgag tgtctcatgg
     7981 gcagataaag cagataatcc gtattcttag caaggcactt gagagttgct taaaaggacc
     8041 tgacacttac aacagtcaag ttctgataga agctacagta atagcactaa ccaaattaca
     8101 gccacttctt aataaggact cgcctctgca caaagccctc ttttgggtag ctgtggctgt
     8161 gctgcagctt gatgaggtca acttgtattc agcaggtacc gcacttcttg aacaaaacct
     8221 gcatacttta gatagtctcc gtatattcaa tgacaagagt ccagaggaag tatttatggc
     8281 aatccggaat cctctggagt ggcactgcaa gcaaatggat cattttgttg gactcaattt
     8341 caactctaac tttaactttg cattggttgg acacctttta aaagggtaca ggcatccttc
     8401 acctgctatt gttgcaagaa cagtcagaat tttacataca ctactaactc tggttaacaa
     8461 acacagaaat tgtgacaaat ttgaagtgaa tacacagagc gtggcctact tagcagcttt
     8521 acttacagtg tctgaagaag ttcgaagtcg ctgcagccta aaacatagaa agtcacttct
     8581 tcttactgat atttcaatgg aaaatgttcc tatggataca tatcccattc atcatggtga
     8641 cccttcctat aggacactaa aggagactca gccatggtcc tctcccaaag gttctgaagg
     8701 ataccttgca gccacctatc caactgtcgg ccagaccagt ccccgagcca ggaaatccat
     8761 gagcctggac atggggcaac cttctcaggc caacactaag aagttgcttg gaacaaggaa
     8821 aagttttgat cacttgatat cagacacaaa ggctcctaaa aggcaagaaa tggaatcagg
     8881 gatcacaaca ccccccaaaa tgaggagagt agcagaaact gattatgaaa tggaaactca
     8941 gaggatttcc tcatcacaac agcacccaca tttacgtaaa gtttcagtgt ctgaatcaaa
     9001 tgttctcttg gatgaagaag tacttactga tccgaagatc caggcgctgc ttcttactgt
     9061 tctagctaca ctggtaaaat ataccacaga tgagtttgat caacgaattc tttatgaata
     9121 cttagcagag gccagtgttg tgtttcccaa agtctttcct gttgtgcata atttgttgga
     9181 ctctaagatc aacaccctgt tatcattgtg ccaagatcca aatttgttaa atccaatcca
     9241 tggaattgtg cagagtgtgg tgtaccatga agaatcccca ccacaatacc aaacatctta
     9301 cctgcaaagt tttggtttta atggcttgtg gcggtttgca ggaccgtttt caaagcaaac
     9361 acaaattcca gactatgctg agcttattgt taagtttctt gatgccttga ttgacacgta
     9421 cctgcctgga attgatgaag aaaccagtga agaatccctc ctgactccca catctcctta
     9481 ccctcctgca ctgcagagcc agcttagtat cactgccaac cttaaccttt ctaattccat
     9541 gacctcactt gcaacttccc agcattcccc aggaatcgac aaggagaacg ttgaactctc
     9601 ccctaccact ggccactgta acagtggacg aactcgccac ggatccgcaa gccaagtgca
     9661 gaagcaaaga agcgctggca gtttcaaacg taatagcatt aagaagatcg tgtagtaatg
     9721 aactagtccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     9781 cgacgcaaca ttgagggtag tccatccacg cgttaagtcg acaatcaacc tctggattac
     9841 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     9901 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     9961 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
    10021 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
    10081 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
    10141 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
    10201 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
    10261 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
    10321 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
    10381 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
    10441 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
    10501 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
    10561 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
    10621 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
    10681 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
    10741 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
    10801 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
    10861 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    10921 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
    10981 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
    11041 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
    11101 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
    11161 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
    11221 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
    11281 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
    11341 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
    11401 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
    11461 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
    11521 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
    11581 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
    11641 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
    11701 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
    11761 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
    11821 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
    11881 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
    11941 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
    12001 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
    12061 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
    12121 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
    12181 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
    12241 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
    12301 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
    12361 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
    12421 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
    12481 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
    12541 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
    12601 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
    12661 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
    12721 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
    12781 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
    12841 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
    12901 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    12961 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    13021 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
    13081 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    13141 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    13201 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    13261 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    13321 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    13381 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    13441 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    13501 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    13561 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    13621 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    13681 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
    13741 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    13801 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    13861 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
    13921 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
    13981 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
    14041 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
    14101 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
    14161 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
    14221 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
    14281 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
    14341 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
    14401 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
    14461 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
    14521 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
    14581 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
    14641 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
    14701 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
    14761 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
    14821 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
    14881 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
    14941 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
    15001 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
    15061 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
    15121 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
    15181 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
    15241 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
    15301 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
    15361 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
    15421 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
    15481 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
    15541 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
    15601 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
    15661 cccaaccccg aggggaccca tgcattgcat ctcaattagt cagcaaccag gtgtggaaag
    15721 tccccaggct ccccagcagg cagaagtatg caaagcatgc gtctcaatta gtcagcaacc
    15781 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
    15841 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
    15901 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
    15961 tctagaggta ccaccggagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
    16021 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
    16081 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
    16141 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
    16201 cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
    16261 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
    16321 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
    16381 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
    16441 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
    16501 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
    16561 catgacccgc aagcccggtg ccagttaacg aattctagat cttgagacaa atggcagtat
    16621 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
    16681 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
    16741 aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggcct
    16801 gcaggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt
    16861 ctaggtcttg aaaggagtgg gaatt
//
