LOCUS       sequence-394871-       14295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Big-IN Payload yeast assembly vector. Encodes LEU2 yeast selectable 
            marker and components for bacterial copy number induction. Contains 
            a mammalian transient (backbone) GFP-T2A-BSD selection cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14295)
  AUTHORS   Brosh R, Coelho C, Ribeiro-Dos-Santos AM, Ellis G, Hogan MS, Ashe 
            HJ, Somogyi N, Ordonez R, Luther RD, Huang E, Boeke JD, Maurano MT
  TITLE     Synthetic regulatory genomics uncovers enhancer context dependence 
            at the Sox2 locus.
  JOURNAL   Mol Cell. 2023 Apr 6;83(7):1140-1152.e7. doi: 
            10.1016/j.molcel.2023.02.027. Epub 2023 Mar 16.
  PUBMED    36931273
REFERENCE   2  (bases 1 to 14295)
  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.molcel.2023.02.027"; journalName: "Mol Cell"; date: 
            "2023-04-6- 6"; volume: "83"; issue: "7"; pages: "1140-1152.e7"
COMMENT     Sequence Label: pRB051_LEU2
FEATURES             Location/Qualifiers
     source          1..14295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(88..765)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             772..783
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    801..817
                     /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)."
     promoter        complement(join(825..831,832..849,850..855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    870..891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(1003..1036)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1517..2611
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(3188..4003)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3188..4003)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3911..3930
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(4004..4106)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     mobile_element  4134..4901
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     misc_feature    4903..5406
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     rep_origin      6257..6871
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      6348..6879
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6947..7166
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             7257..8012
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    8015..8265
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             8591..9766
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             9766..10737
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    10810..11283
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     promoter        11460..12638
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          11691..12629
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     12586..12606
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(12659..12661,12662..12664,12665..13375)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(12659..12661,12662..12664,12665..13375)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(12704..12725)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(12965..12984)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     13312..13333
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             13385..13438
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             13439..13837
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(13865..13882)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13871..14078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ataacttcgt ataggatact ttatacgaag ttatcagtag agacggcctg gctctagtag
       61 cgatctacac tagcactatc agcgttatta agcaccggtg gagtgacgac cttcagcacg
      121 ttcgtactgt tcaacgatgg tgtagtcttc gttgtgggag gtgatgtcca gtttgatgtc
      181 ggttttgtaa gcacccggca gctgaaccgg ttttttagcc atgtaggtgg ttttaacttc
      241 agcgtcgtag tgaccaccgt ctttcagttt cagacgcatt ttgatttcac ctttcagagc
      301 accgtcttcc gggtacatac gttcggtgga agcttcccaa cccatggttt ttttctgcat
      361 aaccggaccg tcggacggga agttggtacc acgcagttta actttgtaga tgaactcacc
      421 gtcttgcagg gaggagtcct gggtaacggt aacaacacca ccgtcttcga agttcataac
      481 acgttcccat ttgaaacctt ccgggaagga cagtttcagg tagtccggga tgtcagccgg
      541 gtgtttaacg taagctttgg aaccgtactg gaactgcggg gacaggatgt cccaagcgaa
      601 cggcagcgga ccacctttgg taactttcag tttagcggtc tgggtacctt cgtacggacg
      661 accttcacct tcaccttcga tttcgaactc gtgaccgtta acggaacctt ccatacgaac
      721 tttgaaacgc atgaactctt tgataacgtc ttcggaggaa gccatctagt atttctcctc
      781 tttctctagt atgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      841 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      901 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      961 caacgcgcgg ggagaggcgg tttgcgtatt gcgtctcatt ttataacttc gtatagcata
     1021 cattatacga agttattagg gataacaggg taattttagg gtagcatcag gaatctgaac
     1081 cctcagaaag tggggatccc gggtatagac ctttatctgc ggttcaagtt aggcataagg
     1141 ctgcatgcta ccttgtcaca cctacactgc tcgaagtaaa tatgggaagc gtgcgacctg
     1201 gctccaggcg ttccgcgccg ccacgtgttc gttaactgtt gattggtggc acatagaggt
     1261 cgcctgacgc atataccttt ttcaactgaa aaattgggag aaaaaggaaa ggtgagagcg
     1321 ccggaaccgg cttttcatat agaatagaga agcgttcatg actaaatgct tgcatcacaa
     1381 tacttgaagt tgacaatatt atttaaggac ctattgtttt ttccaatagg tggttagcaa
     1441 tcgtcttact ttctaacttt tcttaccttt tacatttcag caatatatat atatatttca
     1501 aggatatacc attctaatgt ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt
     1561 tggtcaagaa atcacagccg aagccattaa ggttcttaaa gctatttctg atgttcgttc
     1621 caatgtcaag ttcgatttcg aaaatcattt aattggtggt gctgctatcg atgctacagg
     1681 tgttccactt ccagatgagg cgctggaagc ctccaagaag gctgatgccg ttttgttagg
     1741 tgctgtgggt ggtcctaaat ggggtaccgg tagtgttaga cctgaacaag gtttactaaa
     1801 aatccgtaaa gaacttcaat tgtacgccaa cttaagacca tgtaactttg catccgactc
     1861 tcttttagac ttatctccaa tcaagccaca atttgctaaa ggtactgact tcgttgttgt
     1921 cagagaatta gtgggaggta tttactttgg taagagaaag gaagacgatg gtgatggtgt
     1981 cgcttgggat agtgaacaat acaccgttcc agaagtgcaa agaatcacaa gaatggccgc
     2041 tttcatggcc ctacaacatg agccaccatt gcctatttgg tccttggata aagctaatgt
     2101 tttggcctct tcaagattat ggagaaaaac tgtggaggaa accatcaaga acgaattccc
     2161 tacattgaag gttcaacatc aattgattga ttctgccgcc atgatcctag ttaagaaccc
     2221 aacccaccta aatggtatta taatcaccag caacatgttt ggtgatatca tctccgatga
     2281 agcctccgtt atcccaggtt ccttgggttt gttgccatct gcgtccttgg cctctttgcc
     2341 agacaagaac accgcatttg gtttgtacga accatgccac ggttctgctc cagatttgcc
     2401 aaagaataag gtcaacccta tcgccactat cttgtctgct gcaatgatgt tgaaattgtc
     2461 attgaacttg cctgaagaag gtaaggccat tgaagatgca gttaaaaagg ttttggatgc
     2521 aggtatcaga actggtgatt taggtggttc caacagtacc accgaagtcg gtgatgctgt
     2581 cgccgaagaa gttaagaaaa tccttgctta aaaagattct ctttttttat gatatttgta
     2641 cataaacttt ataaatgaaa ttcataatag aaacgacacg aaattacaaa atggaatatg
     2701 ttcatagggt agacgaaact atatacgcaa tctacataca tttatcaaga aggagaaaaa
     2761 ggaggatgta aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa
     2821 aaagaattgc actttaacat taatattgac aaggaggagg gcaccgcggc cgcccaacta
     2881 cgcagccgat caacaacaga aacgcgacgg tcgagatgag gaacgcacac tggagccttt
     2941 atcagatggt gttgatttca attactccgg agcgtgatgc ggatcaaagt aaccgcgatg
     3001 gaaggtgggg gactcgaaat agatccgttc ttagccctgc gaaacatcta ctgtacacca
     3061 catgcaacag ccacaattct catgtttgac agcttatcat cgaatttctg ccattcatcc
     3121 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
     3181 aaaaaaatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     3241 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     3301 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     3361 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     3421 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     3481 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     3541 gtgattgcgc ctgagccaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     3601 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     3661 caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg gtgagtaacc
     3721 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     3781 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     3841 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     3901 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     3961 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cattttagct tccttagctc
     4021 ctgaaaatct cgataactca aaaaatacgt ccggtagtga tcttatttca ttatggtgaa
     4081 agttggaacc tcttacgtgc cgatcaacgt ccctgggtcc ttttcatcac gtgggtgatg
     4141 ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt
     4201 tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaaagcacc
     4261 gccggacatc agcgctatct ctgctctcac tgccgtaaaa catggcaact gcagttcact
     4321 tacaccgctt ctcaacccgg tacgcaccag aaaatcattg atatggccat gaatggcgtt
     4381 ggatgccggg caacagcccg cattatgggc gttggcctca acacgatttt acgtcactta
     4441 aaaaactcag gccgcagtcg gtaacctcgc gcatacagcc gggcagtgac gtcatcgtct
     4501 gcgcggaaat ggacgaacag tggggctatg tcggggctaa atcgcgccag cgctggctgt
     4561 tttacgcgta tgacagtctc cggaagacgg ttgttgcgca cgtattcggt gaacgcacta
     4621 tggcgacgct ggggcgtctt atgagcctgc tgtcaccctt tgacgtggtg atatggatga
     4681 cggatggctg gccgctgtat gaatcccgcc tgaagggaaa gctgcacgta atcagcaagc
     4741 gatatacgca gcgaattgag cggcataacc tgaatctgag gcagcacctg gcacggctgg
     4801 gacggaagtc gctgtcgttc tcaaaatcgg tggagctgca tgacaaagtc atcgggcatt
     4861 atctgaacat aaaacactat caataagttg gagtcattac ccatcacgtg ctataaaaat
     4921 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     4981 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     5041 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     5101 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     5161 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     5221 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     5281 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     5341 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     5401 gcgatctttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
     5461 tatttattct gcgaagtgat cttccgtcac aggtatttat tcgcgataag ctcatggagc
     5521 ggcgtaaccg tcgcacagga aggacagaga aagcgcggat ctgggaagtg acggacagaa
     5581 cggtcaggac ctggattggg gaggcggttg ccgccgctgc tgctgacggt gtgacgttct
     5641 ctgttccggt cacaccacat acgttccgcc attcctatgc gatgcacatg ctgtatgccg
     5701 gtataccgct gaaagttctg caaagcctga tgggacataa gtccatcagt tcaacggaag
     5761 tctacacgaa ggtttttgcg ctggatgtgg ctgcccggca ccgggtgcag tttgcgatgc
     5821 cggagtctga tgcggttgcg atgctgaaac aattatcctg agaataaatg ccttggcctt
     5881 tatatggaaa tgtggaactg agtggatatg ctgtttttgt ctgttaaaca gagaagctgg
     5941 ctgttatcca ctgagaagcg agcgaaacag tcgggaaaat ctcccattat cgtagagatc
     6001 cgcattatta atctcaggag cctgtgtagc gtttatagga agtagtgttc tgtcatgatg
     6061 cctgcaagcg gtaacgaaaa cgatttgaat atgccttcag gaacaataga aatcttcgtg
     6121 cggtgttacg ttgaagtgga gcggattatg tcagcaatgg acagaacaac ctaatgaaca
     6181 cagaaccatg atgtggtctg tccttttaca gccagtagtg ctcgccgcag tcgagcgaca
     6241 gggcgaagcc ctcgagctgg ttgccctcgc cgctgggctg gcggctgtct atggccctgc
     6301 aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac cgcggccggc cgccggcgtt
     6361 gtggatacct cgcggaaaac ttggccctca ctgacagatg aggggcggac gttgacactt
     6421 gaggggccga ctcacccggc gcggcgttga cagatgaggg gcaggctcga tttcggccgg
     6481 cgacgtggag ctggacagcc tcgcaaatcg gcgaaaacgc ctgattttac gcgagtttcc
     6541 cacagatgat gtggacaagc ctggggataa gtgccctgcg gtattgacac ttgaggggcg
     6601 cgactactga cagatgaggg gcgcgatcct tgacacttga ggggcagagt gctgacagat
     6661 gaggggcgca cctattgaca tttgaggggc tgtccacagg cagaaaatcc agcatttgca
     6721 agggtttccg cccgtttttc ggccaccgct aacctgtctt ttaacctgct tttaaaccaa
     6781 tatttataaa ccttgttttt aaccagggct gcgccctgtg cgcgtgaccg cgcacgccga
     6841 aggggggtgc ccccccttct cgaaccctcc cggtcgagtg agcgaggaag caccagggaa
     6901 cagcacttat atattctgct tacacacgat gcctgaaaaa acttcccttg gggttatcca
     6961 cttatccacg gggatatttt tataattatt ttttttatag tttttagatc ttctttttta
     7021 gagcgccttg taggccttta tccatgctgg ttctagagaa ggtgttgtga caaattgccc
     7081 tttcagtgtg acaaatcacc ctcaaatgac agtcctgtct gtgacaaatt gcccttaacc
     7141 ctgtgacaaa ttgccctcag aagaagctgt tttttcacaa agttatccct gcttattgac
     7201 tcttttttat ttagtgtgac aatctaaaaa cttgtcacac ttcacatgga tctgtcatgg
     7261 cggaaacagc ggttatcaat cacaagaaac gtaaaaatag cccgcgaatc gtccagtcaa
     7321 acgacctcac tgaggcggca tatagtctct cccgggatca aaaacgtatg ctgtatctgt
     7381 tcgttgacca gatcagaaaa tctgatggca ccctacagga acatgacggt atctgcgaga
     7441 tccatgttgc taaatatgct gaaatattcg gattgacctc tgcggaagcc agtaaggata
     7501 tacggcaggc attgaagagt ttcgcgggga aggaagtggt tttttatcgc cctgaagagg
     7561 atgccggcga tgaaaaaggc tatgaatctt ttccttggtt tatcaaacgt gcgcacagtc
     7621 catccagagg gctttacagt gtacatatca acccatatct cattcccttc tttatcgggt
     7681 tacagaaccg gtttacgcag tttcggctta gtgaaacaaa agaaatcacc aatccgtatg
     7741 ccatgcgttt atacgaatcc ctgtgtcagt atcgtaagcc ggatggctca ggcattgtct
     7801 ctctgaaaat cgactggatc atagagcgtt accagctgcc tcaaagttac cagcgtatgc
     7861 ctgacttccg ccgccgcttc ctgcaggtct gtgttaatga gatcaacagc agaactccaa
     7921 tgcgcctctc atacattgag aaaaagaaag gccgccagac gactcatatc gtattttcct
     7981 tccgcgatat cacttccatg acgacaggat agtctgaggg ttatctgtca cagatttgag
     8041 ggtggttcgt cacatttgtt ctgacctact gagggtaatt tgtcacagtt ttgctgtttc
     8101 cttcagcctg catggatttt ctcatacttt ttgaactgta atttttaagg aagccaaatt
     8161 tgagggcagt ttgtcacagt tgatttcctt ctctttccct tcgtcatgtg acctgatatc
     8221 gggggttagt tcgtcatcat tgatgagggt tgattatcac agtttattac tctgaattgg
     8281 ctatccgcgt gtgtacctct acctggagtt tttcccacgg tggatatttc ttcttgcgct
     8341 gagcgtaaga gctatctgac agaacagttc ttctttgctt cctcgccagt tcgctcgcta
     8401 tgctcggtta cacggctgcg gcgagcgcta gtgataataa gtgactgagg tatgtgctct
     8461 tcttatctcc ttttgtagtg ttgctcttat tttaaacaac tttgcggttt tttgatgact
     8521 ttgcgatttt gttgttgctt tgcagtaaat tgcaagattt aataaaaaaa cgcaaagcaa
     8581 tgattaaagg atgttcagaa tgaaactcat ggaaacactt aaccagtgca taaacgctgg
     8641 tcatgaaatg acgaaggcta tcgccattgc acagtttaat gatgacagcc cggaagcgag
     8701 gaaaataacc cggcgctgga gaataggtga agcagcggat ttagttgggg tttcttctca
     8761 ggctatcaga gatgccgaga aagcagggcg actaccgcac ccggatatgg aaattcgagg
     8821 acgggttgag caacgtgttg gttatacaat tgaacaaatt aatcatatgc gtgatgtgtt
     8881 tggtacgcga ttgcgacgtg ctgaagacgt atttccaccg gtgatcgggg ttgctgccca
     8941 taaaggtggc gtttacaaaa cctcagtttc tgttcatctt gctcaggatc tggctctgaa
     9001 ggggctacgt gttttgctcg tggaaggtaa cgacccccag ggaacagcct caatgtatca
     9061 cggatgggta ccagatcttc atattcatgc agaagacact ctcctgcctt tctatcttgg
     9121 ggaaaaggac gatgtcactt atgcaataaa gcccacttgc tggccggggc ttgacattat
     9181 tccttcctgt ctggctctgc accgtattga aactgagtta atgggcaaat ttgatgaagg
     9241 taaactgccc accgatccac acctgatgct ccgactggcc attgaaactg ttgctcatga
     9301 ctatgatgtc atagttattg acagcgcgcc taacctgggt atcggcacga ttaatgtcgt
     9361 atgtgctgct gatgtgctga ttgttcccac gcctgctgag ttgtttgact acacctccgc
     9421 actgcagttt ttcgatatgc ttcgtgatct gctcaagaac gttgatctta aagggttcga
     9481 gcctgatgta cgtattttgc ttaccaaata cagcaatagc aatggctctc agtccccgtg
     9541 gatggaggag caaattcggg atgcctgggg aagcatggtt ctaaaaaatg ttgtacgtga
     9601 aacggatgaa gttggtaaag gtcagatccg gatgagaact gtttttgaac aggccattga
     9661 tcaacgctct tcaactggtg cctggagaaa tgctctttct atttgggaac ctgtctgcaa
     9721 tgaaattttc gatcgtctga ttaaaccacg ctgggagatt agataatgaa gcgtgcgcct
     9781 gttattccaa aacatacgct caatactcaa ccggttgaag atacttcgtt atcgacacca
     9841 gctgccccga tggtggattc gttaattgcg cgcgtaggag taatggctcg cggtaatgcc
     9901 attactttgc ctgtatgtgg tcgggatgtg aagtttactc ttgaagtgct ccggggtgat
     9961 agtgttgaga agacctctcg ggtatggtca ggtaatgaac gtgaccagga gctgcttact
    10021 gaggacgcac tggatgatct catcccttct tttctactga ctggtcaaca gacaccggcg
    10081 ttcggtcgaa gagtatctgg tgtcatagaa attgccgatg ggagtcgccg tcgtaaagct
    10141 gctgcactta ccgaaagtga ttatcgtgtt ctggttggcg agctggatga tgagcagatg
    10201 gctgcattat ccagattggg taacgattat cgcccaacaa gtgcttatga acgtggtcag
    10261 cgttatgcaa gccgattgca gaatgaattt gctggaaata tttctgcgct ggctgatgcg
    10321 gaaaatattt cacgtaagat tattacccgc tgtatcaaca ccgccaaatt gcctaaatca
    10381 gttgttgctc ttttttctca ccccggtgaa ctatctgccc ggtcaggtga tgcacttcaa
    10441 aaagccttta cagataaaga ggaattactt aagcagcagg catctaacct tcatgagcag
    10501 aaaaaagctg gggtgatatt tgaagctgaa gaagttatca ctcttttaac ttctgtgctt
    10561 aaaacgtcat ctgcatcaag aactagttta agctcacgac atcagtttgc tcctggagcg
    10621 acagtattgt ataagggcga taaaatggtg cttaacctgg acaggtcccg tgttccaact
    10681 gagtgtatag agaaaattga ggccattctt aaggaacttg aaaagccagc accctgatgc
    10741 gacctcgttt tagtctacgt ttatctgtct ttacttaatg tcctttgtta caggccagaa
    10801 agcataactg gcctgaatat tctctctggg cccactgttc cacttgtatc gtcggtctga
    10861 taatcagact gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca
    10921 cggtcccact cgtatcgtcg gtctgattat tagtctggga ccacggtccc actcgtatcg
    10981 tcggtctgat aatcagactg ggaccacggt cccactcgta tcgtcggtct gattattagt
    11041 ctgggaccat ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacggtccca
    11101 ctcgtatcgt cggtctgatt attagtctgg aaccacggtc ccactcgtat cgtcggtctg
    11161 attattagtc tgggaccacg gtcccactcg tatcgtcggt ctgattatta gtctgggacc
    11221 acgatcccac tcgtgttgtc ggtctgatta tcggtctggg accacggtcc cacttgtatt
    11281 gtcgatcaga ctatcagcgt gagactacga ttccatcaat gcctgtcaag ggcaagtatt
    11341 gacatgtcgt cgtaacctgt agaacggagt aacctcggtg tgcggttgta tgcctgcggt
    11401 ggattgctgc tgtgtcctgc ttatccacaa cattttgcgc acggttatgt ggacgggccg
    11461 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
    11521 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
    11581 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
    11641 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
    11701 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
    11761 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
    11821 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
    11881 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
    11941 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
    12001 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
    12061 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
    12121 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgccta
    12181 gtctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
    12241 ttgcgtgagc ggaaagatgg ccgcttcccg gccctactgc agggagctca aaatggagga
    12301 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
    12361 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gctgtccagg cacctcgatt
    12421 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
    12481 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
    12541 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
    12601 tggttcaaag tttttttctt ccatttcagg tgtcgtgatc tctagaagct gggtaccaat
    12661 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
    12721 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
    12781 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
    12841 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
    12901 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
    12961 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
    13021 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
    13081 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
    13141 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
    13201 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
    13261 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
    13321 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcag
    13381 tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccaat
    13441 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
    13501 cagcatcccc atctctgagg actacagcgt cgccagcgca gctctctcta gcgacggccg
    13561 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
    13621 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
    13681 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
    13741 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
    13801 tgaattgctg ccctctggtt atgtgtggga gggctaagaa ttctaactag agctcgctga
    13861 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
    13921 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
    13981 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
    14041 ggggaggatt gggaagagaa tagcaggcat gctggggata aggtactacc aatatttagt
    14101 ttctaagcct tgcgacagac ctcccactta gattgccacg catagagcta gcgagtcagc
    14161 gaaaagcatg acgcgctttc aagcgtggcg agtatgtgaa ccaaggcttc ggacaggact
    14221 atatacttag gtttgatctc gccccgagaa ctgtaaacct caacatttat agattattag
    14281 ggataacagg gtaat
//
