LOCUS       sequence-396964-       10114 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 10114)
  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 10114)
  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: TFORF2402
FEATURES             Location/Qualifiers
     source          1..10114
                     /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    5971..6559
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6024..6044)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6442..6453)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6631..6864
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6942..7063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6979..6998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7033..7052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7103..7238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7258..7277)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7259..7277)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7287..7304)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7287..7303)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7296..7318)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7445..7900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7532..7551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7742..7763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7926..8030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8031..8099,8100..8891)
                     /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(8249..8268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9062..9650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9551..9570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9804..9821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9938..9959
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9974..9979,9980..9997,9998..10004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10012..10028
                     /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     10017..10039
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10036..10052
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10036..10052
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10071..10091
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10073..10091
                     /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 caccatggat ttgattgaca tactttggag
     4141 gcaagatata gatcttggag taagtcgaga agtatttgac ttcagtcagc gacggaaaga
     4201 gtatgagctg gaaaaacaga aaaaacttga aaaggaaaga caagaacaac tccaaaagga
     4261 gcaagagaaa gcctttttcg ctcagttaca actagatgaa gagacaggtg aatttctccc
     4321 aattcagcca gcccagcaca tccagtcaga aaccagtgga tctgccaact actcccaggt
     4381 tgcccacatt cccaaatcag atgctttgta ctttgatgac tgcatgcagc ttttggcgca
     4441 gacattcccg tttgtagatg acaatgagtc acttgttcct gatattcccg gtcacatcga
     4501 gagcccagtc ttcattgcta ctaatcaggc tcagtcacct gaaacttctg ttgctcaggt
     4561 agcccctgtt gatttagacg gtatgcaaca ggacattgag caagtttggg aggagctatt
     4621 atccattcct gagttacagt gtcttaatat tgaaaatgac aagctggttg agactaccat
     4681 ggttccaagt ccagaagcca aactgacaga agttgacaat tatcattttt actcatctat
     4741 accctcaatg gaaaaagaag taggtaactg tagtccacat tttcttaatg cttttgagga
     4801 ttccttcagc agcatcctct ccacagaaga ccccaaccag ttgacagtga actcattaaa
     4861 ttcagatgcc acagtcaaca cagattttgg tgatgaattt tattctgctt tcatagctga
     4921 gcccagtatc agcaacagca tgccctcacc tgctacttta agccattcac tctctgaact
     4981 tctaaatggg cccattgatg tttctgatct atcactttgc aaagctttca accaaaacca
     5041 ccctgaaagc acagcagaat tcaatgattc tgactccggc atttcactaa acacaagtcc
     5101 cagtgtggca tcaccagaac actcagtgga atcttccagc tatggagaca cactacttgg
     5161 cctcagtgat tctgaagtgg aagagctaga tagtgcccct ggaagtgtca aacagaatgg
     5221 tcctaaaaca ccagtacatt cttctgggga tatggtacaa cccttgtcac catctcaggg
     5281 gcagagcact cacgtgcatg atgcccaatg tgagaacaca ccagagaaag aattgcctgt
     5341 aagtcctggt catcggaaaa ccccattcac aaaagacaaa cattcaagcc gcttggaggc
     5401 tcatctcaca agagatgaac ttagggcaaa agctctccat atcccattcc ctgtagaaaa
     5461 aatcattaac ctccctgttg ttgacttcaa cgaaatgatg tccaaagagc agttcaatga
     5521 agctcaactt gcattaattc gggatatacg taggaggggt aagaataaag tggctgctca
     5581 gaattgcaga aaaagaaaac tggaaaatat agtagaacta gagcaagatt tagatcattt
     5641 gaaagatgaa aaagaaaaat tgctcaaaga aaaaggagaa aatgacaaaa gccttcacct
     5701 actgaaaaaa caactcagca ccttatatct cgaagttttc agcatgctac gtgatgaaga
     5761 tggaaaacct tattctccta gtgaatactc cctgcagcaa acaagagatg gcaatgtttt
     5821 ccttgttccc aaaagtaaga agccagatgt taagaaaaac tagtaatgaa ctagtccgta
     5881 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg agatgtgtga
     5941 aagatgctat cgaggacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa
     6001 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     6061 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     6121 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6181 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6241 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6301 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6361 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6421 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6481 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6541 ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc
     6601 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     6661 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6721 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6781 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6841 agtcagtgtg gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt
     6901 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     7081 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     7141 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     7201 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     7261 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     7321 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     7381 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     7441 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     7501 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     7561 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     7621 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     7681 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7741 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7801 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7861 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     7921 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7981 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     8041 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     8101 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     8161 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     8221 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     8281 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     8341 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     8401 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     8461 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     8521 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     8581 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     8641 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     8701 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     8761 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     8821 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     8881 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     8941 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     9001 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     9061 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     9121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     9181 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     9241 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     9301 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     9361 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     9421 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     9481 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     9541 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     9601 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     9661 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     9721 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     9781 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     9841 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     9901 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     9961 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
    10021 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
    10081 ctcactaaag ggaacaaaag ctggagctgc aagc
//
