LOCUS       sequence-391512-        9406 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 9406)
  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 9406)
  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: TFORF0507
FEATURES             Location/Qualifiers
     source          1..9406
                     /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    5263..5851
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5316..5336)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5734..5745)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5923..6156
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6234..6355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6271..6290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6325..6344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6395..6530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6550..6569)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6551..6569)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6579..6596)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6579..6595)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6588..6610)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6737..7192
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6824..6843)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7034..7055
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7218..7322
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7323..7391,7392..8183)
                     /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(7541..7560)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8354..8942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8843..8862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9096..9113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9230..9251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9266..9271,9272..9289,9290..9296)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9304..9320
                     /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     9309..9331
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9328..9344
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9328..9344
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9363..9383
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9365..9383
                     /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 caccatggag aacttccaaa aggtggaaaa
     4141 gatcggagag ggcacgtacg gagttgtgta caaagccaga aacaagttga cgggagaggt
     4201 ggtggcgctt aagaaaatcc gcctggacac tgagactgag ggtgtgccca gtactgccat
     4261 ccgagagatc tctctgctta aggagcttaa ccatcctaat attgtcaagc tgctggatgt
     4321 cattcacaca gaaaataaac tctacctggt ttttgaattt ctgcaccaag atctcaagaa
     4381 attcatggat gcctctgctc tcactggcat tcctcttccc ctcatcaaga gctatctgtt
     4441 ccagctgctc cagggcctag ctttctgcca ttctcatcgg gtcctccacc gagaccttaa
     4501 acctcagaat ctgcttatta acacagaggg ggccatcaag ctagcagact ttggactagc
     4561 cagagctttt ggagtccctg ttcgtactta cacccatgag gtggtgaccc tgtggtaccg
     4621 agctcctgaa atcctcctgg gctgcaaata ttattccaca gctgtggaca tctggagcct
     4681 gggctgcatc tttgctgaga tgcacctagt gggtacccag caccatgcta ggtgctgtgg
     4741 ggaacacaga agaaatggaa gacagagtct ctgcccgctg tgctcgtatc tagaagtggc
     4801 tgcatcacaa ggttggggga tgaccgcagt gtctaccccc taccccgtga ctcgccgggc
     4861 cctattccct ggagattctg agattgacca gctcttccgg atctttcgga ctctggggac
     4921 cccagatgag gtggtgtggc caggagttac ttctatgcct gattacaagc caagtttccc
     4981 caagtgggcc cggcaagatt ttagtaaagt tgtacctccc ctggatgaag atggacggag
     5041 cttgttatcg caaatgctgc actacgaccc taacaagcgg atttcggcca aggcagccct
     5101 ggctcaccct ttcttccagg atgtgaccaa gccagtaccc catcttcgac tctagtaatg
     5161 aactagtccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     5221 cgagatactt gctcgaacat ctctatcacg cgttaagtcg acaatcaacc tctggattac
     5281 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     5341 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     5401 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     5461 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     5521 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     5581 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     5641 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     5701 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     5761 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     5821 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     5881 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5941 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6001 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6061 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6121 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     6181 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     6241 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6301 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6361 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6421 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6481 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     6541 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     6601 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     6661 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     6721 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6781 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6841 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6901 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6961 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     7021 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     7081 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     7141 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     7201 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7261 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7321 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     7381 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     7441 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     7501 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7561 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7621 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7681 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7741 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7801 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7861 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7921 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7981 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     8041 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     8101 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     8161 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     8221 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     8281 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     8341 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     8401 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     8461 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     8521 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8581 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8641 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8701 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8761 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8821 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8881 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8941 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     9001 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     9061 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     9121 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     9181 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     9241 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     9301 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     9361 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagc
//
