LOCUS       sequence-404579-       10804 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 10804)
  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 10804)
  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: TFORF1058
FEATURES             Location/Qualifiers
     source          1..10804
                     /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    6661..7249
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6714..6734)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7132..7143)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7321..7554
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7632..7753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7669..7688)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7723..7742
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7793..7928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7948..7967)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7949..7967)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7977..7994)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7977..7993)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7986..8008)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8135..8590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8222..8241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8432..8453
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8616..8720
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8721..8789,8790..9581)
                     /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(8939..8958)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9752..10340
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10241..10260
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10494..10511
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10628..10649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10664..10669,10670..10687,10688..10694)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10702..10718
                     /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     10707..10729
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10726..10742
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10726..10742
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10761..10781
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10763..10781
                     /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 caccatgacg gggctggagg accaggagtt
     4141 cgacttcgag ttcctcttcg agtttaacca gcgcgacgag ggcgccgccg cggccgcccc
     4201 agaacactat ggctatgcat cctccaacgt cagccccgcc ctgccgctcc ccacggcgca
     4261 ctccaccctg ccggccccgt gccacaacct tcagacctcc acaccgggca tcatcccgcc
     4321 ggcggatcac ccctcggggt acggagcagc tttggacggt gggcccgcgg gctacttcct
     4381 ctcctccggc cacaccaggc ctgatggggc ccctgccctg gagagtcctc gcatcgagat
     4441 aacctcgtgc ttgggcctgt accacaacaa taaccagttt ttccacgatg tggaggtgga
     4501 agacgtcctc cctagctcca aacggtcccc ctccacggcc acgctgagtc tgcccagcct
     4561 ggaggcctac agagacccct cgtgcctgag cccggccagc agcctgtcct cccggagctg
     4621 caactcagag gcctcctcct acgagtccaa ctactcgtac ccgtacgcgt ccccccagac
     4681 gtcgccatgg cagtctccct gcgtgtctcc caagaccacg gaccccgagg agggctttcc
     4741 ccgcgggctg ggggcctgca cactgctggg ttccccgcgg cactccccct ccacctcgcc
     4801 ccgcgccagc gtcactgagg agagctggct gggtgcccgc tcctccagac ccgcgtcccc
     4861 ttgcaacaag aggaagtaca gcctcaacgg ccggcagccg ccctactcac cccaccactc
     4921 gcccacgccg tccccgcacg gctccccgcg ggtcagcgtg accgacgact cgtggttggg
     4981 caacaccacc cagtacacca gctcggccat cgtggccgcc atcaacgcgc tgaccaccga
     5041 cagcagcctg gacctgggag atggcgtccc tgtcaagtcc cgcaagacca ccctggagca
     5101 gccgccctca gtggcgctca aggtggagcc cgtcggggag gacctgggca gccccccgcc
     5161 cccggccgac ttcgcgcccg aagactactc ctctttccag cacatcagga agggcggctt
     5221 ctgcgaccag tacctggcgg tgccgcagca cccctaccag tgggcgaagc ccaagcccct
     5281 gtcccctacg tcctacatga gcccgaccct gcccgccctg gactggcagc tgccgtccca
     5341 ctcaggcccg tatgagcttc ggattgaggt gcagcccaag tcccaccacc gagcccacta
     5401 cgagacggag ggcagccggg gggccgtgaa ggcgtcggcc ggaggacacc ccatcgtgca
     5461 gctgcatggc tacttggaga atgagccgct gatgctgcag cttttcattg ggacggcgga
     5521 cgaccgcctg ctgcgcccgc acgccttcta ccaggtgcac cgcatcacag ggaagaccgt
     5581 gtccaccacc agccacgagg ccatcctctc caacaccaaa gtcctggaga tcccactcct
     5641 gccggagaac agcatgcgag ccgtcattga ctgtgccgga atcctgaaac tcagaaactc
     5701 cgacattgaa cttcggaaag gagagacgga catcgggagg aagaacacac gggtacggct
     5761 ggtgttccgc gttcacgtcc cgcaacccag cggccgcacg ctgtccctgc aggtggcctc
     5821 caaccccatc gaatgctccc agcgctcagc tcaggagctg cctctggtgg agaagcagag
     5881 cacggacagc tatccggtcg tgggcgggaa gaagatggtc ctgtctggcc acaacttcct
     5941 gcaggactcc aaggtcattt tcgtggagaa agccccagat ggccaccatg tctgggagat
     6001 ggaagcgaaa actgaccggg acctgtgcaa gccgaattct ctggtggttg agatcccgcc
     6061 atttcggaat cagaggataa ccagccccgt tcacgtcagt ttctacgtct gcaacgggaa
     6121 gagaaagcga agccagtacc agcgtttcac ctaccttccc gccaacgttc caattataaa
     6181 aacagaaccc actgatgatt atgagcctgc tccaacctgt ggaccggtga gccaggggtt
     6241 aagtcctctc ccaagaccat actacagcca gcagctcgcg atgccacccg accccagctc
     6301 ctgcctcgtg gccggcttcc cgccctgtcc gcagagaagc accctgatgc cagcggcccc
     6361 tggcgtgagc cccaagctcc acgacctttc tcccgctgcc tacaccaagg gcgttgccag
     6421 cccgggccac tgtcacctcg gactcccgca gccggccgga gaggcccccg ccgtccagga
     6481 cgtgcccagg ccagtggcca cgcaccccgg ctcgcccggg cagccacccc cggccctgct
     6541 gccacagcag tagtaatgaa ctagtccgta acttgaaagt atttcgattt cttggcttta
     6601 tatatcttgt ggaaaggacg agttagataa acggccgttc gctagacgcg ttaagtcgac
     6661 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     6721 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     6781 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     6841 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     6901 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     6961 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     7021 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     7081 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     7141 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     7201 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     7261 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     7321 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     7381 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7441 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7501 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt
     7561 atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     7621 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7681 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     7741 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     7801 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     7861 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7921 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     7981 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     8041 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     8101 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     8161 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     8221 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     8281 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     8341 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     8401 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     8461 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     8521 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     8581 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     8641 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     8701 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     8761 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     8821 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     8881 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     8941 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     9001 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     9061 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     9121 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     9181 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     9241 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     9301 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     9361 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     9421 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     9481 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     9541 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     9601 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     9661 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     9721 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     9781 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     9841 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     9901 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     9961 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
    10021 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
    10081 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
    10141 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
    10201 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
    10261 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
    10321 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
    10381 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
    10441 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
    10501 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
    10561 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
    10621 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
    10681 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
    10741 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
    10801 aagc
//
