LOCUS Exported 7831 bp ds-DNA circular SYN 02-MAY-2016 DEFINITION Ligation of Fragment 1 into Fragment 2. ACCESSION . VERSION . KEYWORDS 60432-attachment_U-YH4agBCEoU SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7831) AUTHORS . TITLE Direct Submission JOURNAL Exported Monday, May 2, 2016 from SnapGene 3.1.2 http://www.snapgene.com COMMENT PstI: 5082, 1150, 1092, 506 ScaI: 4336, 2253, 1241 FEATURES Location/Qualifiers source 1..7831 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature complement(135..441) /note="f1(-)ori" misc_feature complement(656..968) /note="PB5'TR" misc_feature 1067..1378 /note="TRE" misc_feature 1381..1499 /note="CMV min promoter" misc_feature 1539..2111 /note="b-globin intron" misc_feature 2176..2200 /note="attB1" misc_feature 2202..3123 /note="hNanog" misc_feature 3124..3149 /note="attB2" misc_feature 3187..3213 /note="HA" misc_feature 3260..3831 /note="EMCV IRES" misc_feature 3835..4551 /note="Venus" misc_feature 4631..4832 /note="PGK pA" misc_feature 4958..5169 /note="bGH pA" misc_feature 5389..5623 /note="PB3'TR" misc_feature complement(6066..6748) /note="ColE1 ori" misc_feature complement(6846..7505) /note="AmpR" ORIGIN 1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccttaac 661 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt 721 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc 781 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg 841 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat 901 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta 961 tagatatcat aatttaaaca agcaaaacca aattaagggc cagctcattc ctcccactca 1021 tgatctatag atctggatcc gtcgacaacg gcaagcttac atcgagttta ccactcccta 1081 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt 1141 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac 1201 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag 1261 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt 1321 taccactccc tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgaggt 1381 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc 1441 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc 1501 cgcgggactc agacacccgg gatcctgaga acttcagggt gagtttgggg acccttgatt 1561 gttctttctt tttcgctatt gtaaaattca tgttatatgg agggggcaaa gttttcaggg 1621 tgttgtttag aatgggaaga tgtcccttgt atcaccatgg accctcatga taattttgtt 1681 tctttcactt tctactctgt tgacaaccat tgtctcctct tattttcttt tcattttctg 1741 taactttttc gttaaacttt agcttgcatt tgtaacgaat ttttaaattc acttttgttt 1801 atttgtcaga ttgtaagtac tttctctaat cacttttttt tcaaggcaat cagggtatat 1861 tatattgtac ttcagcacag ttttagagaa caattgttat aattaaatga taaggtagaa 1921 tatttctgca tataaattct ggctggcgtg gaaatattct tattggtaga aacaactaca 1981 ccctggtcat catcctgcct ttctctttat ggttacaatg atatacactg tttgagatga 2041 ggataaaata ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat gccttcttct 2101 ctttcctaca gctcctgggc aacgtgctgg ttgttgtgct gtctcatcat tttggcaaag 2161 aattcctcga atcaaacaag tttgtacaaa aaagcaggct tcaccatgag tgtggatcca 2221 gcttgtcccc aaagcttgcc ttgctttgaa gcatccgact gtaaagaatc ttcacctatg 2281 cctgtgattt gtgggcctga agaaaactat ccatccttgc aaatgtcttc tgctgagatg 2341 cctcacacgg agactgtctc tcctcttccc tcctccatgg atctgcttat tcaggacagc 2401 cctgattctt ccaccagtcc caaaggcaaa caacccactt ctgcagagaa tagtgtcgca 2461 aaaaaggaag acaaggtccc agtcaagaaa cagaagacca gaactgtgtt ctcttccacc 2521 cagctgtgtg tactcaatga tagatttcag agacagaaat acctcagcct ccagcagatg 2581 caagaactct ccaacatcct gaacctcagc tacaaacagg tgaagacctg gttccagaac 2641 cagagaatga aatctaagag gtggcagaaa aacaactggc cgaagaatag caatggtgtg 2701 acgcagaagg cctcagcacc tacctacccc agcctctact cttcctacca ccagggatgc 2761 ctggtgaacc cgactgggaa ccttccaatg tggagcaacc agacctggaa caattcaacc 2821 tggagcaacc agacccagaa catccagtcc tggagcaacc actcctggaa cactcagacc 2881 tggtgcaccc aatcctggaa caatcaggcc tggaacagtc ccttctataa ctgtggagag 2941 gaatctctgc agtcctgcat gcagttccag ccaaattctc ctgccagtga cttggaggct 3001 gctttggaag ctgctgggga aggccttaat gtaatacagc agaccactag gtattttagt 3061 actccacaaa ccatggattt attcctaaac tactccatga acatgcaacc tgaagacgtg 3121 tgagacccag ctttcttgta caaagtggtt cgattcgaga tctcacgcgt gactagtgcc 3181 atggcctacc catacgacgt gccagactac gcctagcggc cgctcgataa gcttgatatc 3241 gaattccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa 3301 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg 3361 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc 3421 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct 3481 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac 3541 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc 3601 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta 3661 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg 3721 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaacgtcta ggccccccga 3781 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg 3841 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 3901 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 3961 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 4021 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac 4081 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 4141 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 4201 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 4261 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc 4321 aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac 4381 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 4441 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 4501 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc 4561 cagcttatcg actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg 4621 aatagtcgag aaattgatga tctattaaac aataaagatg tccacatgga agtttttcct 4681 gtcatacttt gttaagaagg gtgagaacag agtacctaca ttttgaatgg aaggattgga 4741 gctacggggg tgggggtggg gtgggattag ataaatgcct gctctttact gaaggctctt 4801 tactattgct ttatgataat gtttcatagt tggatatcat aatttaaaca agcaaaacca 4861 aattaagggc cagctcattc ctcccactca tgatctatag atcctaagct tatcgatgat 4921 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 4981 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 5041 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 5101 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 5161 gctctatggc ttctgaggcg gaaagaacct gcagcccaag cttggcgtaa tcatggtcat 5221 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 5281 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 5341 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatcctt tgttacttta 5401 tagaagaaat tttgagtttt tgtttttttt taataaataa ataaacataa ataaattgtt 5461 tgttgaattt attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat 5521 taataaataa acctcgatat acagaccgat aaaacacatg cgtcaatttt acgcatgatt 5581 atctttaacg tacgtcacaa tatgattatc tttctagggt taaagagctc cagcttttgt 5641 tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg 5701 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 6121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 6181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 6241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 6301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 6361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 6421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 6481 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 6541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 6601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 6661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 7141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 7201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 7261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 7321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 7381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 7441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 7501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 7561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 7621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 7681 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7741 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7801 gttccgcgca catttccccg aaaagtgcca c //