LOCUS       sequence-407874-       10216 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 10216)
  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 10216)
  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: TFORF0229
FEATURES             Location/Qualifiers
     source          1..10216
                     /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    6073..6661
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6126..6146)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6544..6555)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6733..6966
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7044..7165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7081..7100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7135..7154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7205..7340
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7360..7379)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7361..7379)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7389..7406)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7389..7405)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7398..7420)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7547..8002
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7634..7653)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7844..7865
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8028..8132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8133..8201,8202..8993)
                     /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(8351..8370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9164..9752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9653..9672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9906..9923
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10040..10061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10076..10081,10082..10099,10100..10106)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10114..10130
                     /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     10119..10141
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10138..10154
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10138..10154
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10173..10193
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10175..10193
                     /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 caccatggaa ctcgtaacat tcagggatgt
     4141 ggccatagaa ttctcccctg aagagtggaa atgtctggac cctgcccagc agaatttgta
     4201 tagagatgtg atgttggaga actacaggaa cctggtctcc ctggctatat gttctccttt
     4261 cagccaagac ctttcaccag tgcaggggat agaagattca ttccacaaac ttatactgaa
     4321 aagatacgag aaatgtggac atgagaattt acaattaaga aaaggctgta aacgtgtgaa
     4381 tgagtgtaag gtgcagaaag gagttaataa tggagtttac cagtgcttgt caactaccca
     4441 gagcaaaata tttcaatgta atacatgtgt taaagttttt agtaaatttt caaattcaaa
     4501 caaacataag ataagacata ctggagagaa accctttaaa tgtacagaat gtggcagatc
     4561 gttttacatg tcacacctaa ctcaacatac aggaattcat gctggagaga aaccctacaa
     4621 atgtgaaaaa tgtggcaaag cctttaatag gtccacatca cttagtaaac ataagagaat
     4681 tcatactgga gagaaaccct acacatgtga agaatgtggc aaagccttta gacggtccac
     4741 agttctgaac gaacataaga aaattcatac tggagagaaa ccctacaaat gtgaagaatg
     4801 tggcaaagcc tttacaaggt ccacaacact gaatgaacac aagaaaattc atactggaga
     4861 gaaaccctac aaatgtaaag aatgtggcaa agcctttaga tggtccacaa gcctgaatga
     4921 acataagaat attcatactg gagagaaacc ctacaaatgt aaagaatgtg gcaaagcctt
     4981 tagacagtcc aggagcctga atgaacataa aaatattcat actggcgaaa aaccctacac
     5041 atgtgaaaaa tgtggcaaag cttttaacca atcctcaagt cttattatac acaggagcat
     5101 tcattctgaa caaaaacttt acaaatgtga agaatgtggc aaagccttta cttggtcctc
     5161 atcccttaat aaacataaga gaattcatac tggagagaaa ccctacacat gtgaagaatg
     5221 tggcaaagct ttttataggt cctcacacct tgctaaacat aagagaattc atactggaga
     5281 gaaaccctac acgtgcgaag aatgtggcaa agcttttaac caatcctcaa ctcttatatt
     5341 acacaagaga atccattctg ggcaaaaacc ttacaaatgt gaagaatgtg gcaaagcctt
     5401 tacacggtcc acaacactga acgaacataa gaaaattcat actggcgaga aaccctacaa
     5461 atgtgaagaa tgtggcaaag ctttcatatg gtccgcaagc ctgaatgaac ataagaatat
     5521 tcatactgga gagaaaccct acaaatgtaa agaatgtggc aaagctttta accaatcctc
     5581 aggccttatt atacacagga gcattcattc tgaacaaaaa ctttacaaat gtgaagaatg
     5641 tggcaaagcc tttactcggt ccacagccct gaatgaacat aagaaaattc attctggaga
     5701 gaaaccctac aaatgcaaag aatgtggcaa agcctataac ttatcctcaa cccttactaa
     5761 acataagaga attcatactg gagagaaacc cttcacatgt gaagaatgtg gcaaagcctt
     5821 caattggtcc tcatccctta ctaaacataa gataattcat actggagaga aatcctacaa
     5881 atgtgaagaa tgtggcaaag cttttaatcg gccctcaacc cttactgtac acaagcgaat
     5941 tcatactggc aaggaacata gttagtaatg aactagtccg taacttgaaa gtatttcgat
     6001 ttcttggctt tatatatctt gtggaaagga cgatgtgtga cccggcctct tcaaataacg
     6061 cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6121 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6181 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6241 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6301 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6361 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6421 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     6481 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6541 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6601 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6661 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     6721 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     6781 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6841 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6901 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6961 ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     7021 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     7081 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7141 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     7201 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     7261 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     7321 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     7381 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     7441 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     7501 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     7561 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     7621 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     7681 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     7741 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     7801 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     7861 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7921 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7981 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     8041 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     8101 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     8161 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     8221 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     8281 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     8341 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     8401 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     8461 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     8521 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     8581 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     8641 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     8701 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     8761 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     8821 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     8881 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     8941 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     9001 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     9061 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     9121 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     9181 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     9241 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     9301 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     9361 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     9421 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     9481 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     9541 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     9601 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     9661 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     9721 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     9781 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     9841 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     9901 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     9961 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
    10021 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
    10081 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
    10141 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
    10201 agctggagct gcaagc
//
