LOCUS       sequence-395924-       10132 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 10132)
  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 10132)
  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: TFORF2319
FEATURES             Location/Qualifiers
     source          1..10132
                     /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    5989..6577
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6042..6062)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6460..6471)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6649..6882
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6960..7081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6997..7016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7051..7070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7121..7256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7276..7295)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7277..7295)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7305..7322)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7305..7321)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7314..7336)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7463..7918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7550..7569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7760..7781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7944..8048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8049..8117,8118..8909)
                     /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(8267..8286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9080..9668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9569..9588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9822..9839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9956..9977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9992..9997,9998..10015,10016..10022)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10030..10046
                     /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     10035..10057
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10054..10070
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10054..10070
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10089..10109
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10091..10109
                     /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 caccatggac ctcgaggagg cggaagagtt
     4141 taaagaacgc tgtactcagt gtgctgctgt ctcatggggt cttactgatg aaggcaaata
     4201 ttattgcact tcttgccaca atgttacaga gagatatcag gaagttacaa acactgatct
     4261 tattcctaat acccaaataa aagccctcaa ccgggggctt aaaaaaaaaa acaatactga
     4321 aaaaggctgg gattggtatg tgtgtgaagg tttccagtat attctttatc aacaagcaga
     4381 agccttaaag aaccttggag taggcccaga gttaaagaac gatgttttac ataatttttg
     4441 gaagcgctac cttcagaaga gcaagcaggc atattgtaag aacccagttt ataccactgg
     4501 aaggaaacct acggtattag aagataatct aagtcattca gactgggcta gtgagcctga
     4561 gctgctaagt gatgtcagct gtcctccttt tcttgaaagt ggagcggagt ctcagtctga
     4621 catccacact cgaaaacctt tccccgtcag caaagcatca caatcagaaa cgtctgtctg
     4681 ctctggatct ctggatggag ttgaatactc acaacgaaag gagaagggaa tcgtgaagat
     4741 gaccatgcca cagacacttg ccttctgtta tctgtcctta ctttggcaga gagaagcaat
     4801 aacactttca gatcttttga ggtttgttga agaggaccat attccttaca taaatgcttt
     4861 tcagcatttt ccagaacaga tgaaattata tggacgtgac agaggaatct ttggtataga
     4921 gtcttggcct gactacgagg acatctacaa aaaaacagta gaagttggaa catttttaga
     4981 tttgcctcgt tttccagaca taactgaaga ctgctatctt catcccaaca tactgtgtat
     5041 gaaatacttg atggaagtca acctccctga tgaaatgcat agcttaactt gccacgtggt
     5101 aaaaatgact ggaatgggag aagtggattt tctgacattt gatcctatag ctaaaatggc
     5161 aaaaactgtt aagtacgatg tacaagctgt agctatcatt gtggtggtat tgaaactgct
     5221 ctttctattg gatgacagtt tcgagtggtc tttgtctaat cttgctgaaa agcataatga
     5281 aaagaacaaa aaagataagc catggtttga tttcagaaag tggtaccaaa ttatgaagaa
     5341 agcttttgat gagaaaaaac aaaaatggga agaagcaagg gccaagtacc tgtggaaaag
     5401 tgaaaagcca ctctactact catttgtcga caaaccagta gcatataaaa aaagagaaat
     5461 ggtggtgaat ctacagaaac aatttagcac actggtcgag tcaacagcaa ctgctggaaa
     5521 aaaaagccct tcaagttttc agttcaactg gactgaagag gacactgata gaacgtgttt
     5581 ccatggacac agccttcagg gagtcctgaa agagaaaggc caatcactgc tgactaagaa
     5641 ttcattatat tggcttagta cacagaaatt ctgcagatgc tattgtacac atgtgacaac
     5701 ctatgaagaa tcaaattatt ctctgagtta tcagtttata ctaaatctct tctccttcct
     5761 gctcagaata aagacttccc ttctccatga agaagtgagc ttagttgaga agaaactttt
     5821 tgagaaaaaa tacagtgtaa aaagaaagaa atcaagatcc aagaaagtga gacgacatta
     5881 gtaatgaact agtccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     5941 aaaggacgag tcggtcgctc ttcagttgtt cttacgcgtt aagtcgacaa tcaacctctg
     6001 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     6061 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     6121 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6181 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     6241 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     6301 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6361 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     6421 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6481 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6541 cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt
     6601 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     6661 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     6721 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6781 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6841 gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat
     6901 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     6961 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7021 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7081 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     7141 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     7201 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     7261 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     7321 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     7381 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     7441 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     7501 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     7561 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     7621 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     7681 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     7741 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     7801 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     7861 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     7921 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     7981 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     8041 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     8101 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     8161 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     8221 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     8281 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     8341 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     8401 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     8461 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     8521 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     8581 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     8641 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     8701 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     8761 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     8821 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     8881 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     8941 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     9001 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     9061 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     9121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     9181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     9241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     9301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     9361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     9421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     9481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     9541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     9601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     9661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     9721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     9781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     9841 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     9901 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     9961 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
    10021 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
    10081 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gc
//
