LOCUS       sequence-400290-       10078 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 10078)
  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 10078)
  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: TFORF1814
FEATURES             Location/Qualifiers
     source          1..10078
                     /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    5935..6523
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5988..6008)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6406..6417)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6595..6828
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6906..7027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6943..6962)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6997..7016
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7067..7202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7222..7241)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7223..7241)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7251..7268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7251..7267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7260..7282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7409..7864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7496..7515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7706..7727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7890..7994
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7995..8063,8064..8855)
                     /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(8213..8232)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9026..9614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9515..9534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9768..9785
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9902..9923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9938..9943,9944..9961,9962..9968)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9976..9992
                     /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     9981..10003
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10000..10016
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10000..10016
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10035..10055
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10037..10055
                     /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 aacctgaagt ctggagtgta
     4141 tcctctcaag gaagcaagtg gatgccctgg ggctgacagg aatcttctgg tgtactcttt
     4201 ttatgaaaag gggccattga catttaggga tgtggccata gaattttctc tggaggagtg
     4261 gcaatgcctg gacactgctc agcaggattt gtatagaaaa gtgatgttag agaactacag
     4321 aaacctggtc ttcttggcag gtattgctgt ttctaagcca gacctgatca cctgtctaga
     4381 gcaaggaaaa gagccctgga atatgaagag acatgcgatg gtagatcaac ccccagttac
     4441 atattctcat tttgcccaag acctttggcc agagcagggc ataaaagatt ctttccaaga
     4501 agtcatattg agaagatatg gaaaatgtgg acatgaagat ttacagttaa gaacaggctg
     4561 taaaagtgtg gatgagtgta atctgcacaa agaatgttat gatgaactaa accagtgttt
     4621 gacaactacc cagagtgaaa tatttcaata tgataaatat gtgaatgtct tttataaatt
     4681 ttcaaatcca aatatacaaa agataagaca tactggaaag aagcctttca aatgtaaaaa
     4741 atgtgacaaa tcgttttgca tgcttttaca cctaactcaa cataaaagaa ttcatattag
     4801 ggaaaattct taccaatgtg aagaatgtgg taaagtcttt aactggttct caacccttac
     4861 tagacacaga agaattcata ctggagagaa accctacaaa tgtgaacaat gtggcaaagc
     4921 ttttaagcag tcctcaaccc ttactacaca taagataatt catactggag agaaacccta
     4981 cagatgtgaa gaatgtggca aaacctttaa ccggtcctca caccttacta cacataaaag
     5041 aattcatact ggagagaaac cctacagatg tgaagaatgt ggcagagctt ttaaccggtc
     5101 ctcacacctt actacacata agattattca tactggagag aaaccctaca aatgtgaaga
     5161 atgtggcaaa gcttttaacc aatcctcaac ccttactaca cataagataa ttcatgctgg
     5221 agagaaacct tacaaatgtg aagaatgtgg caaagctttt taccgattct cataccttac
     5281 taaacataag ataattcata ctggagagaa attctacaaa tgtgaagaat gtggcaaagg
     5341 ctttaattgg tcctcgaccc ttactaaaca taaaagaatt catactggag agaaacccta
     5401 caaatgtgaa caatgtggca aagcttttaa tgagtcctca aaccttactg cacataagat
     5461 aattcatact ggagagaaac cctacaaatg tgaagaatgt ggcaaagcct ttaaccggtc
     5521 cccaaaactt actgcacata aggtaattca ttctggagag aaaccctaca aatgtgaaga
     5581 atgtggcaaa gcttttaacc aattctcaaa ccttactaaa cataagataa ctcatattgg
     5641 agatacatct tacaaatacc tagaatgtga taaagccttt agccagtctt caactcttac
     5701 taaacataag gtaattcata ctggagagaa accctacaac tgtgaagaat acggcaaagc
     5761 tttcaaccag tcctcaaacc ttattgaaca aagtaattca tactggagag aaaccctaca
     5821 aatgtagtaa tgaactagtc cgtaacttga aagtatttcg atttcttggc tttatatatc
     5881 ttgtggaaag gacgatcgat gttgcgtccg ggctcataga cgcgttaagt cgacaatcaa
     5941 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6001 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6061 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6121 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6181 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6241 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6301 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     6361 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6421 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6481 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     6541 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6601 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     6661 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6721 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6781 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta
     6841 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     6901 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7021 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     7081 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     7141 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     7201 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     7261 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     7321 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     7381 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     7441 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     7501 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     7561 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     7621 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     7681 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     7741 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     7801 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     7861 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     7921 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     7981 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     8041 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     8101 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     8161 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     8221 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     8281 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     8341 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     8401 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     8461 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     8521 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     8581 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     8641 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     8701 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     8761 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     8821 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     8881 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     8941 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     9001 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     9061 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     9121 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     9181 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     9241 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     9301 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     9361 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     9421 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     9481 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     9541 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     9601 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     9661 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     9721 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     9781 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     9841 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     9901 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     9961 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
    10021 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagc
//
