LOCUS       sequence-397258-        9028 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 9028)
  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 9028)
  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: TFORF0084
FEATURES             Location/Qualifiers
     source          1..9028
                     /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"
     misc_feature    4885..5473
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4938..4958)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5356..5367)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5545..5778
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5856..5977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5893..5912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5947..5966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6017..6152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6172..6191)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6173..6191)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6201..6218)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6201..6217)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6210..6232)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6359..6814
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6446..6465)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6656..6677
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6840..6944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6945..7013,7014..7805)
                     /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(7163..7182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7976..8564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8465..8484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8718..8735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8852..8873
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8888..8893,8894..8911,8912..8918)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8926..8942
                     /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     8931..8953
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8950..8966
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8950..8966
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8985..9005
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8987..9005
                     /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 caccatgcgg ggccagggcc gcaaggagag
     4141 tttgtccgat tcccgagacc tggacggctc ctacgaccag ctcacggaca gcgtggacga
     4201 tgaatttgaa ttgtccaccg tgtgtcaccg gcctgagggt ctggagcagc tgcaggagca
     4261 aaccaaattc acgcgcaagg agttgcaggt cctgtaccgg ggcttcaaga acgaatgtcc
     4321 cagcggaatt gtcaatgagg agaacttcaa gcagatttac tcccagttct ttcctcaagg
     4381 agactccagc acctatgcca cttttctctt caatgccttt gacaccaacc atgatggctc
     4441 ggtcagtttt gaggactttg tggctggttt gtccgtgatt cttcggggaa ctgtagatga
     4501 caggcttaat tgggccttca acctgtatga ccttaacaag gacggctgca tcaccaagga
     4561 ggaaatgctt gacatcatga agtccatcta tgacatgatg ggcaagtaca cgtaccctgc
     4621 actccgggag gaggccccaa gggaacacgt ggagagcttc ttccagaaga tggacagaaa
     4681 caaggatggt gtggtgacca ttgaggaatt cattgagtct tgtcaaaagg atgagaacat
     4741 catgaggtcc atgcagctct ttgacaatgt catctagtaa tgaactagtc cgtaacttga
     4801 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaagaa ctgagggcga
     4861 tatacgagca cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     4921 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4981 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5041 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5101 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5161 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     5221 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     5281 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5341 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     5401 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5461 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5521 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     5581 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5641 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5701 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5761 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
     5821 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     5881 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5941 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6001 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6061 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6121 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     6181 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6241 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6301 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     6361 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6421 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6481 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6541 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6601 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6661 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6721 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6781 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     6841 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6901 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6961 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     7021 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7081 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7141 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7201 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7261 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7321 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7381 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7441 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7501 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7561 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7621 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7681 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7741 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7801 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7861 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7921 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7981 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8041 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8101 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8161 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8221 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8281 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8341 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8401 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8461 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8521 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8581 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8641 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8701 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8761 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8821 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8881 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8941 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     9001 aaagggaaca aaagctggag ctgcaagc
//
