LOCUS       sequence-462949-       12985 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 12985)
  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 12985)
  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: TFORF2770
FEATURES             Location/Qualifiers
     source          1..12985
                     /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"
     regulatory      4109..4118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    8842..9430
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8895..8915)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9313..9324)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9502..9735
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9813..9934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9850..9869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9904..9923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9974..10109
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10129..10148)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10130..10148)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10158..10175)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10158..10174)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10167..10189)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10316..10771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10403..10422)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10613..10634
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10797..10901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10902..10970,10971..11762)
                     /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(11120..11139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11933..12521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12422..12441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12675..12692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12809..12830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12845..12850,12851..12868,12869..12875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12883..12899
                     /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     12888..12910
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12907..12923
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12907..12923
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12942..12962
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12944..12962
                     /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 caccatggaa ccggggtgtg acgagttcct
     4141 gccgccaccg gagtgcccgg tttttgagcc tagctgggct gaattccaag acccgcttgg
     4201 ctacattgcg aaaataaggc ccatagcaga gaagtctggc atctgcaaaa tccgcccacc
     4261 cgcggattgg cagcctcctt ttgcagtaga agttgacaat ttcagattta ctcctcgcgt
     4321 ccaaaggcta aatgaactgg aggcccaaac tagagtgaaa ttgaactatt tggatcagat
     4381 tgcaaaattc tgggaaattc aaggctcctc tttaaagatt cccaatgtgg agcggaagat
     4441 cttggacctc tacagcctta gtaagattgt gattgaggaa ggtggctatg aagccatctg
     4501 caaggatcgt cggtgggctc gagttgccca gcgtctccac tacccaccag gcaaaaacat
     4561 tggctccctg ctacgatcac attacgaacg cattatttac ccctatgaaa tgtttcagtc
     4621 tggagccaac catgtgcaat gtaacacaca cccgtttgac aatgaggtaa aagataagga
     4681 atacaagccc cacagcatcc cccttagaca gtctgtgcag ccttcaaagt tcagcagcta
     4741 cagtcgacgg gcaaaaaggc tacagcctga tccagagcct acagaggagg acattgagaa
     4801 gcatccagag ctaaagaagt tacagatata tgggccaggt cccaaaatga tgggcttggg
     4861 ccttatggct aaggataagg ataagactgt gcataagaaa gtcacatgcc ccccaactgt
     4921 tacggtgaag gatgagcaaa gtggaggtgg gaacgtgtca tcaacattgc tcaagcagca
     4981 cttgagccta gagccctgca ctaagacaac catgcaactt cgaaagaatc acagcagtgc
     5041 ccagtttatt gactcatata tttgccaagt atgctcccgt ggggatgaag atgataagct
     5101 tcttttctgt gatggctgtg atgacaatta ccacatcttc tgcttgttac caccccttcc
     5161 tgaaatcccc agaggcatct ggaggtgccc aaaatgtatc ttggcggagt gtaaacagcc
     5221 tcctgaagct tttggatttg aacaggctac ccaggagtac agtttgcaga gttttggtga
     5281 aatggctgat tccttcaagt ccgactactt caacatgcct gtacatatgg tgcctacaga
     5341 acttgtagag aaggaattct ggaggctggt gagcagcatt gaggaagacg tgacagttga
     5401 atatggagct gatattcatt ccaaagaatt tggcagtggc tttcctgtca gcaatagcaa
     5461 acaaaactta tctcctgagg agaaggagta tgcgaccagt ggttggaacc tgaatgtgat
     5521 gccagtgcta gatcagtctg ttctctgtca catcaatgca gacatctcag gcatgaaggt
     5581 gccctggctg tacgtgggca tggttttctc agcattttgt tggcatattg aggatcactg
     5641 gagttactct attaactatc tgcattgggg tgagccgaag acctggtatg gtgtaccctc
     5701 cctggcagca gagcatttgg aggaggtgat gaagatgctg acacctgagc tgtttgatag
     5761 ccagcctgat ctcctacacc agcttgtcac tctcatgaat cccaacactt tgatgtccca
     5821 tggtgtgcca gttgtccgca caaaccagtg tgcaggggag tttgtcatca cttttcctcg
     5881 tgcttaccac agtggtttta accaaggcta caattttgct gaagctgtca acttttgtac
     5941 tgctgactgg ctacctgctg gacgccagtg cattgaacac taccgccggc tccggcgcta
     6001 ttgtgtcttc tcccacgagg agctcatctg caagatggct gccttcccag agacgttgga
     6061 tctcaatcta gcagtagctg tgcacaagga gatgttcatt atggttcagg aggagcgacg
     6121 tctacgaaag gcccttttgg agaagggcgt cacggaggct gagcgagagg cttttgagct
     6181 gctcccagat gatgaacgcc agtgcatcaa gtgcaagacc acgtgcttct tgtcagccct
     6241 ggcctgctac gactgcccag atggccttgt atgcctttcc cacatcaatg acctctgcaa
     6301 gtgctctagt agccgacagt acctccggta tcggtacacc ttggatgagc tccccaccat
     6361 gctgcataaa ctgaagattc gggctgagtc ttttgacacc tgggccaaca aagtgcgagt
     6421 ggccttggag gtggaggatg gccgtaaacg cagctttgaa gagctaaggg cactggagtc
     6481 tgaggctcgt gagaggaggt ttcctaatag tgagctgctt cagcgactga agaactgcct
     6541 gagtgaggtg gaggcttgta ttgctcaagt cctggggctg gtcagtggtc aggtggccag
     6601 gatggacact ccacagctga ccttgactga actccgggtc cttcttgagc agatgggcag
     6661 cctgccctgt gccatgcatc agattgggga tgtcaaggat gtcctggaac aggtggaggc
     6721 ctatcaagct gaggctcgtg aggctctggc cacactgccc tctagtccag ggctattgcg
     6781 gtccctgttg gagagggggc agcagctggg tgtagaggtg cctgaagccc atcagcttca
     6841 gcagcaggtg gagcaggcgc aatggctaga tgaagtgaag caggccctgg ccccttctgc
     6901 tcacaggggc tctctggtca tcatgcaggg gcttttggtt atgggtgcca agatagcctc
     6961 cagcccttct gtggacaagg cccgggctga gctgcaagaa ctactgacca ttgcagagcg
     7021 ctgggaagaa aaggctcatt tctgcctgga ggccaggcag aagcatccac cagccacatt
     7081 ggaagccata attcgtgaga cagaaaacat ccctgttcac ctgcctaaca tccaggctct
     7141 caaagaagct ctgactaagg cacaagcttg gattgctgat gtggatgaga tccaaaatgg
     7201 tgaccactac ccctgtctag atgacttgga gggcctggtg gctgtgggcc gggacctgcc
     7261 tgtggggctg gaagagctga gacagctaga gctgcaggta ttgacagcac attcctggag
     7321 agagaaggcc tccaagacct ttctcaagaa gaattcttgc tacacactgc ttgaggtgct
     7381 ttgcccgtgt gcagacgctg gctcagacag caccaagcgt agccggtgga tggagaaggc
     7441 gctggggttg taccagtgtg acacagagct gctggggctg tctgcacagg acctcagaga
     7501 cccaggctct gtgattgtgg ccttcaagga aggggaacag aaggagaagg agggtatcct
     7561 gcagctgcgt cgcaccaact cagccaagcc cagtccactg gcaccatccc tcatggcctc
     7621 ttctccgact tctatctgtg tgtgtgggca ggtgccagct ggggtgggag ttctgcagtg
     7681 tgacctgtgt caggactggt tccatgggca gtgtgtgtca gtgccccatc tcctcacctc
     7741 tccaaagccc agtctcactt catctccact gctagcctgg tgggaatggg acacaaaatt
     7801 cctgtgtcca ctgtgtatgc gctcacgacg gccacgccta gagacaatcc tagccttgct
     7861 ggttgccctg cagaggctgc ccgtgcggct gcctgagggt gaggcccttc agtgtctcac
     7921 agagagggcc attggctggc aagaccgtgc cagaaaggct ctggcctctg aagatgtgac
     7981 tgctctgttg cgacagctgg ctgagcttcg ccaacagcta caggccaaac ccagaccaga
     8041 ggaggcctca gtctacactt cagccactgc ctgtgaccct atcagagaag gcagtggcaa
     8101 caatatttct aaggtccaag ggctgctgga gaatggagac agtgtgacca gtcctgagaa
     8161 catggctcca ggaaagggct ctgacctgga gctactgtcc tcactgttgc cgcagttgac
     8221 tggccctgtg ttggagctgc ctgaggcaat ccgggctccc ctggaggagc tcatgatgga
     8281 aggggacctg cttgaggtga ccctggatga gaaccacagc atctggcagc tgctgcaggc
     8341 tggacagcct ccagacctgg acagaattcg cacacttctg gagctggaaa aatttgaaca
     8401 tcaagggagt cggacaagga gccgggctct ggagaggcga cggcggcggc agaaggtgga
     8461 tcagggtaga aacgttgaga atcttgttca acaggagctt cagtcaaaaa gggctcggag
     8521 ctcagggatt atgtctcagg tgggccgaga agaagaacat tatcaggaga aagcagaccg
     8581 tgaaaatatg ttcctgacac cttccacaga ccacagccct ttcttgaaag gaaaccaaaa
     8641 tagcttacaa cacaaggatt caggctcttc agctgcttgt ccttctttaa tgcctttgct
     8701 acaactctcc tactctgatg agcaacagtt gtagtaatga actagtccgt aacttgaaag
     8761 tatttcgatt tcttggcttt atatatcttg tggaaaggac gatttgagac tcatattgga
     8821 ttaattacgc gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     8881 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     8941 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     9001 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     9061 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     9121 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     9181 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     9241 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     9301 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     9361 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     9421 gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact
     9481 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     9541 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     9601 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     9661 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     9721 ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac
     9781 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     9841 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     9901 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     9961 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    10021 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
    10081 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
    10141 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
    10201 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
    10261 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
    10321 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
    10381 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
    10441 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    10501 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
    10561 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    10621 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
    10681 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
    10741 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
    10801 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
    10861 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
    10921 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
    10981 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
    11041 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
    11101 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
    11161 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    11221 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
    11281 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
    11341 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
    11401 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
    11461 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
    11521 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
    11581 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
    11641 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
    11701 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
    11761 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    11821 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    11881 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    11941 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    12001 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    12061 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
    12121 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    12181 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    12241 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    12301 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    12361 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    12421 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    12481 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    12541 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
    12601 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
    12661 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
    12721 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
    12781 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
    12841 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
    12901 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
    12961 gggaacaaaa gctggagctg caagc
//
