LOCUS       sequence-395859-       10903 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 10903)
  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 10903)
  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: TFORF1465
FEATURES             Location/Qualifiers
     source          1..10903
                     /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    6760..7348
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6813..6833)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7231..7242)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7420..7653
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7731..7852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7768..7787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7822..7841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7892..8027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8047..8066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8048..8066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8076..8093)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8076..8092)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8085..8107)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8234..8689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8321..8340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8531..8552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8715..8819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8820..8888,8889..9680)
                     /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(9038..9057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9851..10439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10340..10359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10593..10610
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10727..10748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10763..10768,10769..10786,10787..10793)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10801..10817
                     /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     10806..10828
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10825..10841
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10825..10841
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10860..10880
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10862..10880
                     /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 ctgtggcgcc aatgcaccca
     4141 ctggctcatc cagtgccggg tgctgccgcc cagccaccgc gtgacctggg atggggctca
     4201 ggtgtgtgaa ctggcccagg ccctccggga tggtgtcctt ctgtgtcagc tgcttaacaa
     4261 cctgctaccc catgccatca acctgcgtga ggtcaacctg cgcccccaga tgtcccagtt
     4321 cctgtgcctt aagaacatta gaaccttcct gtccacctgc tgtgagaagt tcggcctcaa
     4381 gcggagcgag ctcttcgaag cctttgacct cttcgatgtg caggattttg gcaaggtcat
     4441 ctacaccctg tctgctctgt cctggacccc gatcgcccag aacaggggga tcatgccctt
     4501 ccccaccgag gaggagagtg taggtgatga agacatctac agtggcctgt ccgaccagat
     4561 cgacgacacg gtggaggagg atgaggacct gtatgactgc gtggagaatg aggaggcgga
     4621 aggcgacgag atctatgagg acctcatgcg ctcggagccc gtgtccatgc cgcccaagat
     4681 gacagagtat gacaagcgct gctgctgcct gcgggagatc cagcagacgg aggagaagta
     4741 cactgacacg ctgggctcca tccagcagca tttcttgaag cccctgcaac ggttcctgaa
     4801 acctcaagac attgagatca tctttatcaa cattgaggac ctgcttcgtg ttcatactca
     4861 cttcctaaag gagatgaagg aagccctggg cacccctggc gcagccaatc tctaccaggt
     4921 cttcatcaaa tacaaggaga ggttcctcgt ctatggccgc tactgcagcc aggtggagtc
     4981 agccagcaaa cacctggacc gtgtggccgc agcccgggag gacgtgcaga tgaagctgga
     5041 ggaatgttct cagagagcca acaacgggag gttcaccctg cgggacctgc tgatggtgcc
     5101 tatgcagcga gttctcaaat atcacctcct tctccaggag ctggtgaaac acacgcagga
     5161 ggcgatggag aaggagaacc tgcggctggc cctggatgcc atgagggacc tggctcagtg
     5221 cgtgaacgag gtcaagcgag acaacgagac actgcgacag atcaccaatt tccagctgtc
     5281 cattgagaac ctggaccagt ctctggctca ctatggccgg cccaagatcg acggggaact
     5341 caagatcacc tcggtggaac ggcgctccaa gatggacagg tatgccttcc tgctcgacaa
     5401 agctctactc atctgtaagc gcaggggaga ctcctatgac ctcaaggact ttgtaaacct
     5461 gcacagcttc caggttcggg atgactcttc aggagaccga gacaacaaga agtggagcca
     5521 catgttcctc ctgatcgagg accaaggtgc ccagggctat gagctgttct tcaagacaag
     5581 agaattgaag aagaagtgga tggagcagtt tgagatggcc atctccaaca tctatccgga
     5641 gaatgccacc gccaacgggc atgacttcca gatgttctcc tttgaggaga ccacatcctg
     5701 caaggcctgt cagatgctgc ttagaggtac cttctatcag ggctaccgct gccatcggtg
     5761 ccgggcatct gcacacaagg agtgtctggg gagggtccct ccatgtggcc gacatgggca
     5821 agatttccca ggaactatga agaaggacaa actacatcgc agggctcagg acaaaaagag
     5881 gaatgagctg ggtctgccca agatggaggt gtttcaggaa tactacgggc ttcctccacc
     5941 ccctggagcc attggaccct ttctacggct caaccctgga gacattgtgg agctcacgaa
     6001 ggctgaggct gaacagaact ggtgggaggg cagaaataca tctactaatg aaattggctg
     6061 gtttccttgt aacagggtga agccctatgt ccatggccct cctcaggacc tgtctgttca
     6121 tctctggtac gcaggcccca tggagcgggc aggggcagag agcatcctgg ccaaccgctc
     6181 ggacgggact ttcttggtgc ggcagagggt gaaggatgca gcagaatttg ccatcagcat
     6241 taaatataac gtcgaggtca agcacattaa aatcatgaca gcagaaggac tgtaccggat
     6301 cacagagaaa aaggctttcc gggggcttac ggagctggtg gagttttacc agcagaactc
     6361 tctaaaggat tgcttcaagt ctctggacac caccttgcag ttccccttca aggagcctga
     6421 aaagagaacc atcagcaggc cagcagtggg aagcacaaag tattttggca cagccaaagc
     6481 ccgctatgac ttctgcgccc gagaccgatc agagctgtcg ctcaaggagg gtgacatcat
     6541 caagatcctt aacaagaagg gacagcaagg ctggtggcga ggggagatct atggccgggt
     6601 tggctggttc cctgccaact acgtggagga agattattct gaatactgct agtaatgaac
     6661 tagtccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     6721 tgaaattcat gcggaacgtg gtgcacgcgt taagtcgaca atcaacctct ggattacaaa
     6781 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     6841 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     6901 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     6961 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     7021 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     7081 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     7141 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     7201 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     7261 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     7321 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
     7381 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     7441 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     7501 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     7561 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     7621 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta
     7681 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     7741 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     7801 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     7861 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     7921 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     7981 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
     8041 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     8101 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     8161 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     8221 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     8281 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     8341 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     8401 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     8461 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     8521 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     8581 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     8641 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     8701 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     8761 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     8821 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     8881 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     8941 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     9001 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     9061 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     9121 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     9181 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     9241 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     9301 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     9361 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     9421 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     9481 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     9541 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     9601 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     9661 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     9721 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     9781 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     9841 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     9901 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     9961 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
    10021 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
    10081 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
    10141 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
    10201 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
    10261 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
    10321 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
    10381 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
    10441 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
    10501 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
    10561 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
    10621 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
    10681 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
    10741 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
    10801 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
    10861 caattaaccc tcactaaagg gaacaaaagc tggagctgca agc
//
