LOCUS       sequence-396293-       10210 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 10210)
  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 10210)
  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: TFORF2109
FEATURES             Location/Qualifiers
     source          1..10210
                     /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    6067..6655
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6120..6140)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6538..6549)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6727..6960
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7038..7159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7075..7094)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7129..7148
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7199..7334
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7354..7373)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7355..7373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7383..7400)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7383..7399)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7392..7414)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7541..7996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7628..7647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7838..7859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8022..8126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8127..8195,8196..8987)
                     /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(8345..8364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9158..9746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9647..9666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9900..9917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10034..10055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10070..10075,10076..10093,10094..10100)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10108..10124
                     /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     10113..10135
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10132..10148
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10132..10148
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10167..10187
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10169..10187
                     /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 caccatgaaa cagcccaaca ggaaaaggaa
     4141 gcttaatatg gattccaaag agcgtttgga tcaggacgga cgcttggagc aagctgaaga
     4201 ggaaaagaag cccaaggata gcacaacccc tttgagtcac gtcccttcag cggctgcaca
     4261 gggagcatgg tcttgggagt ggtacttgaa agaacagaag gctgtcgcag cacctgttga
     4321 gctgttttcc aaggatcagt cctttccaga gcatgaaaat ggttttcaga ttggaatgag
     4381 attagaaggc attgatcccc gacatccatc ggtattctgt gtgctttctg tagcggaggt
     4441 ttgtggttac cgtctaagac ttcattttga tggttattta agttgctatg atttttggac
     4501 caatgctggt tcccctgaca ttcatccagt aggatggtgt gaaaagacca aacatgaact
     4561 gcacatccct aagggttata gaaaagataa atttgtttgg atggattact tgaaggcctg
     4621 caaattgcaa aatgctccaa agaaattatt cagaaacaga agtcctaatg ggccaatgtc
     4681 taaagaattt caggttggaa tgaagctgga ggccgtggac aggaagaacc cttccttggt
     4741 gtgtgtggcg accatagcag atattgttga agatcgctta ctagtgcatt ttgacaactg
     4801 ggatgatagt tacgattact ggtgcgatgt taatagccct tatgtccagc cagttggttg
     4861 gtgtcaggag aatggaagaa ctctgatagc accccaaggt tatcccaatc cagaaaattt
     4921 ttcctggaca gaatacctgg aagctactca aaccaatgca gttcctgcca aagtttttaa
     4981 aatgaggttg cctcatggtt ttctgccaaa tatgaaactt gaagttgtgg ataaacggaa
     5041 ccccaggtta attcgtgttg ctacgattgt agatgttgat gaccaaagag taaaggttca
     5101 ttttgatggt tgggaccata agtatgacta ctgggtggag gcagacagcc ctgatatcca
     5161 cccgatcgga tggtgtgatg tcacagggca tccactggaa gtgccacagc gaacgaatga
     5221 cctgaagatc cttccaggtc aagctgtctg tcctactccc gggtgccgag gaataggcca
     5281 tatccgtggt ccacgttatt cgggacatca cagtgctttt ggctgcccgt attcagacat
     5341 gaacttgaaa aaggaggcaa cacttcacga tcgtttgaga gaacaaacac aggcaaattt
     5401 ggaatcagac tcttcacatt caaaatcaaa aagcctctgt agtttgaact tcaatggaaa
     5461 acatgaaaag gtgaacagtc agcccagact tgtacagcag gcaaagtgtt tgaaaatcaa
     5521 aggaaaagaa gatattgact tggataatct cttcagagaa tactcggtgg agcaggcgca
     5581 gcaggtgctt caccagtcag tgtccatgtc cacggtgtca gcccaccctt ttcgggacct
     5641 tcccctcggc agggagcaac actgcaagtt gcttccaggc gtggctgaca tccgggccag
     5701 ccaagtggca cgttggactg tggatgaggt ggctgagttt gtacagtctc ttctgggctg
     5761 tgaagagcat gccaagtgct ttaagaaaga gcagatcgat ggcaaagcct tcctgcttct
     5821 gacacagacg gacattgtca aagtgatgaa gatcaaactg ggcccagcac tgaagattta
     5881 caactctatc ctgatgttca ggcattccca ggaactccct gaagaagata ttgcctcagg
     5941 ccaagaagtc aggggatagt aatgaactag tccgtaactt gaaagtattt cgatttcttg
     6001 gctttatata tcttgtggaa aggacgagct aacccgactc ttcggtataa cacgcgttaa
     6061 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     6121 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     6181 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     6241 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     6301 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     6361 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     6421 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     6481 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     6541 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6601 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
     6661 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6721 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     6781 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6841 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     6901 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     6961 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     7021 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     7081 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     7141 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     7201 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     7261 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     7321 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     7381 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     7441 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     7501 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     7561 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     7621 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7681 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     7741 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     7801 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7861 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     7921 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     7981 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     8041 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     8101 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     8161 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     8221 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     8281 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     8341 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     8401 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     8461 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     8521 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     8581 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     8641 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     8701 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     8761 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     8821 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     8881 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     8941 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     9001 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     9061 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     9121 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     9181 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     9241 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     9301 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     9361 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     9421 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     9481 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     9541 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     9601 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     9661 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     9721 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     9781 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     9841 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     9901 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     9961 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
    10021 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
    10081 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
    10141 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
    10201 agctgcaagc
//
