LOCUS       sequence-285185-       12394 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 12394)
  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 12394)
  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: TFORF0331
FEATURES             Location/Qualifiers
     source          1..12394
                     /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    8251..8839
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8304..8324)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8722..8733)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8911..9144
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9222..9343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9259..9278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9313..9332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9383..9518
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9538..9557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9539..9557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9567..9584)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9567..9583)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9576..9598)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9725..10180
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9812..9831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10022..10043
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10206..10310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10311..10379,10380..11171)
                     /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(10529..10548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11342..11930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11831..11850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12084..12101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12218..12239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12254..12259,12260..12277,12278..12284)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12292..12308
                     /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     12297..12319
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12316..12332
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12316..12332
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12351..12371
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12353..12371
                     /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 gagaacgagg tggagagcag
     4141 cagcgacgcg gcccctgggc ctggccggcc cgaggagccc tctgagagcg gcctgggtgt
     4201 gggcacctca gaagccgtgt ccgccgacag cagcgacgcc gcggccgccc cggggcaggc
     4261 agaggccgat gactctggcg tggggcaaag ctcggaccgc ggcagccgtt ctcaggagga
     4321 ggtatctgag agcagctcga gcgcagaccc cctgcctaat agctacctcc ctgattcatc
     4381 gtctgtgtct catgggccag tggcaggggt gacaggcggt cccccagcac ttgtgcactc
     4441 tagtgcactc ccagacccca acatgctggt gtccgactgc acagcttcct cctcggacct
     4501 gggctcggcc atcgacaaga tcatcgagtc caccatcggg cccgacctca tccagaactg
     4561 catcactgtg accagtgctg aggatggcgg ggccgagacc acacggtacc tgatcctaca
     4621 gggcccagat gatggagccc ccatgacatc accaatgtcc agttccacct tggcccacag
     4681 cctagcagcc attgaggccc tggcagatgg ccccacatcc acatccacat gcctggaggc
     4741 acagggtggg cccagctccc cggtgcagct gcccccagcc tccggtgccg aagagccgga
     4801 cctgcagagc ctggaggcca tgatggaggt ggtggtggtg cagcagttca aatgcaagat
     4861 gtgccagtac cggagcagca ccaaggccac actgctgcgc cacatgcggg aacgccactt
     4921 ccgtccagta gcagcagccg cagcagcagc tggtaaaaaa ggacgtctac ggaagtggag
     4981 cacctccacc aagagccaag aggaagaggg accagaggag gaggacgatg atgacattgt
     5041 agacgctgga gccattgatg acctggagga ggatagcgac tataatccag ctgaggatga
     5101 gccccgaggc cggcagcttc ggctccagcg ccccaccccc agtaccccaa ggccccgaag
     5161 gagacctggc cggccccgga agctgccccg cctggagatc tcagacctcc cagatggtgt
     5221 ggaaggagag cctctagtga gttcccagag tggacagagc cctccagagc cacaggatcc
     5281 cgaggctccc agctcctcag gcccaggaca cctggtggcc atgggcaagg tgagcaggac
     5341 ccctgtggaa gctggtgtga gccagtcaga tgcagagaac gcagccccct cctgcccgga
     5401 tgagcatgac actctgcccc ggcgccgagg tcgaccttcc aggcgcttcc taggcaagaa
     5461 ataccgcaag tactattaca agtcgcccaa accacttttg aggcccttcc tgtgccgcat
     5521 ctgtggttct cgctttctgt cccacgagga cctgcgcttc cacgtcaact cccatgaggc
     5581 tggcgatccc cagctcttca agtgcctgca gtgcagctat cgttcccgcc gctggtcctc
     5641 gctcaaggag cacatgttca accacgtggg cagcaagccc tacaagtgtg acgagtgcag
     5701 ctacaccagt gtctaccgga aggacgtcat tcggcacgcc gctgtgcaca gccgggaccg
     5761 gaagaagagg ccagatccga ctccaaagct gagctctttc ccctgccctg tgtgtggccg
     5821 tgtgtacccc atgcagaaaa gactcacgca gcacatgaag acgcacagca ctgagaagcc
     5881 ccacatgtgt gacaagtgtg gaaagtcctt taagaagcgc tacaccttca aaatgcacct
     5941 gctcacgcac atccaggctg ttgccaaccg caggttcaag tgtgagttct gtgagtttgt
     6001 ttgtgaagac aagaaggcac tgctgaacca ccagttgtcc cacgtcagtg acaagccctt
     6061 caaatgcagc ttttgtccct accgcacctt ccgagaggac ttcttgctgt cccatgtggc
     6121 tgtcaagcac acaggggcca agcccttcgc ctgtgagtac tgccacttca gcacacggca
     6181 caagaagaac ctgcgcctgc acgtacggtg ccgacacgca agcagcttcg aggaatgggg
     6241 gaggcgccac cctgaggagc ccccctcccg ccgtcgcccc ttcttctctc tgcagcagat
     6301 tgaggagctg aagcagcagc acagtgcggc ccctggacca cctcccagtt ccccaggacc
     6361 tcctgagata cccccagagg cgacaacttt ccagtcatct gaggctccct cattgctctg
     6421 ttctgacacc ctgggcggcg ccaccatcat ctaccagcaa ggagctgagg agtcgacagc
     6481 gatggccacg cagacagcct tggatcttct gctgaacatg agtgctcagc gggaactggg
     6541 gggcacagcc ctgcaggtgg ctgtggtgaa gtcggaagat gtggaagcag ggttagcatc
     6601 ccctggtggg cagccctccc ctgaaggtgc cactccacag gtggtcaccc tccacgtggc
     6661 agagccaggg ggcggtgcag cagccgagag ccagctaggc cctcctgacc taccgcagat
     6721 caccctggca cctggtccat ttggtgggac tggctacagt gtcatcacag caccccctat
     6781 ggaggaggga acatcagctc ctggcacacc ttacagcgag gagcccgcag gagaggcagc
     6841 ccaggctgtg gttgtgagtg acaccctaaa agaagctggc acccactaca tcatggctac
     6901 tgatggtacc cagttgcacc acattgagct caccgcagat ggctccatct ccttcccaag
     6961 tccagatgct ctggcctctg gtgccaaatg gcccctgctg cagtgtgggg gactgcccag
     7021 agacggccct gagcccccat ctccagccaa gacccactgc gtaggggact cccagagctc
     7081 tgcctcctca cctcctgcaa ccagcaaagc cctgggcctg gcagtgcccc cgtcaccgcc
     7141 atctgcagcc actgctgcat caaagaagtt ttcctgcaag atctgtgccg aggccttccc
     7201 tggccgagct gagatggaga gtcacaagcg ggcccacgct gggcctggtg ccttcaagtg
     7261 ccccgactgc cccttcagtg cccgccagtg gcccgaggtc cgggcgcaca tggcacagca
     7321 ctcaagccta cggccccacc agtgtagcca gtgcagcttt gcctccaaga acaagaagga
     7381 cctgcgtcgg cacatgctga ctcacacaaa ggagaagcct tttgcatgcc acctctgcgg
     7441 gcagcgtttc aaccgtaacg ggcacctcaa gttccacatc cagcggctgc acagtcctga
     7501 tgggaggaag tcaggaaccc ctacagcccg ggcccctacc cagaccccaa cccagaccat
     7561 catcctgaac agtgatgacg aaacactggc caccctgcac actgcactcc agtccagtca
     7621 cggggtcctg ggcccagagc ggctacagca ggcactgagc caggaacaca tcatcgttgc
     7681 ccaggaacag acagtgacca atcaggagga agccgcctac atccaagaga tcaccacggc
     7741 agatggccag accgtacagc acctggtgac ctccgacaac caggtgcagt atatcatctc
     7801 ccaggatggt gtccagcacc tgctccccca ggaatatgtt gtggtccctg aaggccatca
     7861 catccaggta caggagggcc agatcacaca catccagtat gaacaaggag ccccgttcct
     7921 tcaggagtcc cagatccagt atgtgcctgt gtccccaggc cagcagcttg tcacacaggc
     7981 tcaacttgag gctgcagcac actcagctgt cacagcagtg gctgatgctg ccatggccca
     8041 agcccagggc ctgtttggta cagacgagac agtgcccgaa cacattcaac agctgcagca
     8101 ccagggcatc gagtacgacg tcatcaccct ggccgatgac tagtaatgaa ctagtccgta
     8161 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaccgtccgg
     8221 tctttattac ccgtcacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa
     8281 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     8341 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     8401 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     8461 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     8521 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     8581 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     8641 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     8701 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     8761 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     8821 ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc
     8881 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     8941 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     9001 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     9061 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     9121 agtcagtgtg gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt
     9181 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     9241 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     9301 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     9361 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     9421 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     9481 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     9541 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     9601 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     9661 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     9721 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     9781 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     9841 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     9901 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     9961 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
    10021 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
    10081 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
    10141 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
    10201 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
    10261 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
    10321 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
    10381 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
    10441 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
    10501 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
    10561 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
    10621 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
    10681 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
    10741 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
    10801 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
    10861 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    10921 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    10981 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    11041 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    11101 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    11161 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    11221 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    11281 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    11341 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    11401 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    11461 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    11521 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    11581 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    11641 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    11701 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    11761 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    11821 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    11881 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    11941 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    12001 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    12061 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
    12121 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
    12181 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
    12241 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
    12301 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
    12361 ctcactaaag ggaacaaaag ctggagctgc aagc
//
