LOCUS       sequence-454958-        9721 bp    DNA     circular SYN 15-DEC-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 9721)
  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 9721)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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: TFORF0478
FEATURES             Location/Qualifiers
     source          1..9721
                     /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    5578..6166
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5631..5651)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6049..6060)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6238..6471
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6549..6670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6586..6605)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6640..6659
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6710..6845
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6865..6884)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6866..6884)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6894..6911)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6894..6910)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6903..6925)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7052..7507
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7139..7158)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7349..7370
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7533..7637
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7638..7706,7707..8498)
                     /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(7856..7875)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8669..9257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9158..9177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9411..9428
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9545..9566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9581..9586,9587..9604,9605..9611)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9619..9635
                     /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     9624..9646
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9643..9659
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9643..9659
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9678..9698
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9680..9698
                     /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 caccatggca aagctctacg aggcagtgac
     4141 cttcaaggat gtggctgtga tcttcactga ggaggagctg gggctgctgg accctgccca
     4201 gaggaagctg taccgagatg tgatgcttga gaacttcagg aacctgctct cagtgggagg
     4261 caagatccaa actgagatgg agactgttcc agaagcagga acacatgaag aattttcctg
     4321 caagcaaatt tgggaacaaa ttgcaagtga cttaaccagg tctcaagata ccaccataag
     4381 taactctcag ttatttgaac aagatgacaa cccctcccag attaaagcaa gactatctac
     4441 agttcacaca agagaaaaac ctttccaggg tgaaaattgt aaacagttct tcagtgatgt
     4501 ttccttcttt gatcttcctc agcagttata ttcaggagag aagtctcata cctgtgatga
     4561 gtgtggaaaa agcttctgtt acatctcagc ccttcatatt catcagaggg tccacatggg
     4621 agtgaaatgc tataagtgtg atgtgtgtgg taaggaattt agtcagagct cacgtctgca
     4681 aactcatcaa agagtccaca ctggagagaa accattcaaa tgtgagcagt gtgggaaagg
     4741 cttcagatgt agatcagcac ttaaagttca ttgcaaatta cacatgagag agaaacctta
     4801 taattgtgag aaatgtggga aggctttcat gcacaatttc cagcttcaga aacatcacag
     4861 aattcatact ggggagaagc cattcaaatg tgaaatatgt ggtaagagct tctgtcttag
     4921 gtcaagtctt aataggcatt gcatggtcca cacagcagag aaactgtaca aatctgaaaa
     4981 gtatggaaga ggtttcattg ataggctaga tttgcataag catcagatga ttcatatggg
     5041 acagaaacca tataattgta aagaatgtgg gaagagcttc aaatggtcct catatctttt
     5101 ggtccatcaa cgagtccaca ctggagaaaa gccatacaaa tgtgaggagt gtgggaaggg
     5161 ctacattagt aagtcaggtc ttgacttcca ccatagaacc cacacgggag agagatctta
     5221 taactgtgat aactgcggga agagctttag acatgcttct agtattttga atcataagaa
     5281 actccactgc caaagaaagc cattgaaatg tgaagactgt ggaaagaggc ttgtatgccg
     5341 gtcatactgt aaagaccaac aaagagacca cagtggagaa aacccatcca aatgtgagga
     5401 ctgtgggaag cgctacaaga ggcgcttgaa tctggatata attttatcat tatttttaaa
     5461 tgacatatag taatgaacta gtccgtaact tgaaagtatt tcgatttctt ggctttatat
     5521 atcttgtgga aaggacgagt caacggacat cgagtgatat tgacgcgtta agtcgacaat
     5581 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     5641 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5701 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5761 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5821 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5881 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5941 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     6001 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     6061 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     6121 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     6181 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     6241 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     6301 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     6361 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     6421 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     6481 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6541 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6601 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6661 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     6721 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6781 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6841 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     6901 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     6961 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     7021 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     7081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     7141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     7201 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     7261 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     7321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     7381 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     7501 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     7561 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     7621 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     7681 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     7741 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     7801 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     7861 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     7921 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     7981 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     8041 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     8101 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     8161 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     8221 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     8281 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     8341 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     8401 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     8461 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     8521 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     8581 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     8641 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     8701 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8761 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     8821 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     8881 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     8941 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     9001 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     9061 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     9121 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     9181 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     9241 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     9301 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     9361 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     9421 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     9481 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     9541 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     9601 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     9661 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
     9721 c
//
