LOCUS       sequence-397809-       10453 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 10453)
  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 10453)
  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: TFORF1837
FEATURES             Location/Qualifiers
     source          1..10453
                     /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    6310..6898
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6363..6383)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6781..6792)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6970..7203
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7281..7402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7318..7337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7372..7391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7442..7577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7597..7616)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7598..7616)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7626..7643)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7626..7642)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7635..7657)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7784..8239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7871..7890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8081..8102
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8265..8369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8370..8438,8439..9230)
                     /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(8588..8607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9401..9989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9890..9909
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10143..10160
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10277..10298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10313..10318,10319..10336,10337..10343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10351..10367
                     /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     10356..10378
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10375..10391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10375..10391
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10410..10430
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10412..10430
                     /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 caccatgggc cagactggga agaaatctga
     4141 gaagggacca gtttgttggc ggaagcgtgt aaaatcagag tacatgcgac tgagacagct
     4201 caagaggttc agacgagctg atgaagtaaa gagtatgttt agttccaatc gtcagaaaat
     4261 tttggaaaga acggaaatct taaaccaaga atggaaacag cgaaggatac agcctgtgca
     4321 catcctgact tcttgttcgg tgaccagtga cttggatttt ccaacacaag tcatcccatt
     4381 aaagactctg aatgcagttg cttcagtacc cataatgtat tcttggtctc ccctacagca
     4441 gaattttatg gtggaagatg aaactgtttt acataacatt ccttatatgg gagatgaagt
     4501 tttagatcag gatggtactt tcattgaaga actaataaaa aattatgatg ggaaagtaca
     4561 cggggataga gaatgtgggt ttataaatga tgaaattttt gtggagttgg tgaatgccct
     4621 tggtcaatat aatgatgatg acgatgatga tgatggagac gatcctgaag aaagagaaga
     4681 aaagcagaaa gatctggagg atcaccgaga tgataaagaa agccgcccac ctcggaaatt
     4741 tccttctgat aaaatttttg aagccatttc ctcaatgttt ccagataagg gcacagcaga
     4801 agaactaaag gaaaaatata aagaactcac cgaacagcag ctcccaggcg cacttcctcc
     4861 tgaatgtacc cccaacatag atggaccaaa tgctaaatct gttcagagag agcaaagctt
     4921 acactccttt catacgcttt tctgtaggcg atgttttaaa tatgactgct tcctacatcc
     4981 ttttcatgca acacccaaca cttataagcg gaagaacaca gaaacagctc tagacaacaa
     5041 accttgtgga ccacagtgtt accagcattt ggagggagca aaggagtttg ctgctgctct
     5101 caccgctgag cggataaaga ccccaccaaa acgtccagga ggccgcagaa gaggacggct
     5161 tcccaataac agtagcaggc ccagcacccc caccattaat gtgctggaat caaaggatac
     5221 agacagtgat agggaagcag ggactgaaac ggggggagag aacaatgata aagaagaaga
     5281 agagaagaaa gatgaaactt cgagctcctc tgaagcaaat tctcggtgtc aaacaccaat
     5341 aaagatgaag ccaaatattg aacctcctga gaatgtggag tggagtggtg ctgaagcctc
     5401 aatgtttaga gtcctcattg gcacttacta tgacaatttc tgtgccattg ctaggttaat
     5461 tgggaccaaa acatgtagac aggtgtatga gtttagagtc aaagaatcta gcatcatagc
     5521 tccagctccc gctgaggatg tggatactcc tccaaggaaa aagaagagga aacaccggtt
     5581 gtgggctgca cactgcagaa agatacagct gaaaaagggt caaaaccgct ttccgggatg
     5641 ccgctgcaaa gcacagtgca acaccaagca gtgcccgtgc tacctggctg tccgagagtg
     5701 tgaccctgac ctctgtctta cttgtggagc cgctgaccat tgggacagta aaaatgtgtc
     5761 ctgcaagaac tgcagtattc agcggggctc caaaaagcat ctattgctgg caccatctga
     5821 cgtggcaggc tgggggattt ttatcaaaga tcctgtgcag aaaaatgaat tcatctcaga
     5881 atactgtgga gagattattt ctcaagatga agctgacaga agagggaaag tgtatgataa
     5941 atacatgtgc agctttctgt tcaacttgaa caatgatttt gtggtggatg caacccgcaa
     6001 gggtaacaaa attcgttttg caaatcattc ggtaaatcca aactgctatg caaaagttat
     6061 gatggttaac ggtgatcaca ggataggtat ttttgccaag agagccatcc agactggcga
     6121 agagctgttt tttgattaca gatacagcca ggctgatgcc ctgaagtatg tcggcatcga
     6181 aagagaaatg gaaatccctt agtaatgaac tagtccgtaa cttgaaagta tttcgatttc
     6241 ttggctttat atatcttgtg gaaaggacga tctgagcaag caacgtgtta gccgacgcgt
     6301 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     6361 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     6421 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     6481 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     6541 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     6601 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     6661 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     6721 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     6781 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     6841 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     6901 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6961 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     7021 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     7081 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     7141 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7201 gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     7261 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     7321 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     7381 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     7441 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     7501 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     7561 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     7621 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     7681 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     7741 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     7801 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     7861 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     7921 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     7981 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     8041 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     8101 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     8161 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     8221 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     8281 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     8341 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     8401 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     8461 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     8521 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     8581 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     8641 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     8701 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     8761 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     8821 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     8881 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     8941 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     9001 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     9061 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     9121 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     9181 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     9241 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     9301 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     9361 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     9421 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     9481 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     9541 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     9601 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     9661 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     9721 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     9781 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     9841 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     9901 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     9961 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    10021 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
    10081 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
    10141 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
    10201 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
    10261 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
    10321 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
    10381 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
    10441 tggagctgca agc
//
