LOCUS Exported 6412 bp ds-DNA circular SYN 24-AUG-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCXB-EBNA1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6412) AUTHORS Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A, Goshima N, Yamanaka S TITLE An Efficient Non-viral Method to Generate Integration-Free Human iPS Cells from Cord Blood and Peripheral Blood Cells. JOURNAL Stem Cells. 2012 Nov 29. doi: 10.1002/stem.1293. PUBMED 23193063 REFERENCE 2 (bases 1 to 6412) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 24, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6412 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(83..102) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 148..203 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(202..221) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(564..580) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(564..580) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(577..599) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 588..604 /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(612..642) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 657..678 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(803..820) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(974..1562) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1054..1073) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1733..2593) /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 2356..2375 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2594..2698) /gene="bla" /label=AmpR promoter enhancer 2729..3108 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3110..3387 /label=chicken beta-actin promoter intron 3388..4404 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 4328..4350 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 4412..4431 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 4474..6399 /codon_start=1 /product="Epstein-Barr nuclear antigen 1, also known as EBNA-1" /label=EBNA1 /note="crucial for latent viral infection, and for episomal amplification of vectors containing the oriP origin (Okita et al., 2013)" /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE GEEGQE" ORIGIN 1 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc 61 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga 121 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt 181 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac 241 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga 301 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct 361 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat 421 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc 481 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc 541 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 601 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 661 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 721 tcgtgccagc ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 781 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 841 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 901 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 961 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 1021 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 1081 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 1141 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 1201 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 1261 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 1321 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 1381 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 1441 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 1501 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 1561 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 1621 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 1681 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 1741 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 1801 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 1861 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 1921 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 1981 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 2041 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 2101 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 2161 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 2221 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 2281 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 2341 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 2401 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 2461 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 2521 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 2581 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 2641 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 2701 catttccccg aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa 2761 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 2821 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 2881 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 2941 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt 3001 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac 3061 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc 3121 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta 3181 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gsgcgcgcca 3241 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc 3301 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc 3361 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc 3421 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag 3481 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg 3541 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc 3601 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc 3661 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc 3721 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag 3781 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg 3841 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct 3901 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg 3961 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag 4021 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc 4081 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag 4141 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc 4201 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt 4261 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc 4321 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt 4381 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat 4441 cattttggca aagaattcgc ccttgtcgac accatgtctg acgaggggcc aggtacagga 4501 cctggaaatg gcctaggaga gaagggagac acatctggac cagaaggctc cggcggcagt 4561 ggacctcaaa gaagaggggg tgataaccat ggacgaggac ggggaagagg acgaggacga 4621 ggaggcggaa gaccaggagc cccgggcggc tcaggatcag ggccaagaca tagagatggt 4681 gtccggagac cccaaaaacg tccaagttgc attggctgca aagggaccca cggtggaaca 4741 ggagcaggag caggagcggg aggggcagga gcaggagggg caggagcagg aggaggggca 4801 ggagcaggag gaggggcagg aggggcagga ggggcaggag gggcaggagc aggaggaggg 4861 gcaggagcag gaggaggggc aggaggggca ggaggggcag gagcaggagg aggggcagga 4921 gcaggaggag gggcaggagg ggcaggagca ggaggagggg caggaggggc aggaggggca 4981 ggagcaggag gaggggcagg agcaggagga ggggcaggag gggcaggagc aggaggaggg 5041 gcaggagggg caggaggggc aggagcagga ggaggggcag gagcaggagg ggcaggaggg 5101 gcaggagggg caggagcagg aggggcagga gcaggaggag gggcaggagg ggcaggaggg 5161 gcaggagcag gaggggcagg agcaggaggg gcaggagcag gaggggcagg agcaggaggg 5221 gcaggagggg caggagcagg aggggcagga ggggcaggag caggaggggc aggaggggca 5281 ggagcaggag gaggggcagg aggggcagga gcaggaggag gggcaggagg ggcaggagca 5341 ggaggggcag gaggggcagg agcaggaggg gcaggagggg caggagcagg aggggcagga 5401 ggggcaggag caggaggagg ggcaggagca ggaggggcag gagcaggagg tggaggccgg 5461 ggtcgaggag gcagtggagg ccggggtcga ggaggtagtg gaggccgggg tcgaggaggt 5521 agtggaggcc gccggggtag aggacgtgaa agagccaggg ggggaagtcg tgaaagagcc 5581 agggggagag gtcgtggacg tggagaaaag aggcccagga gtcccagtag tcagtcatca 5641 tcatccgggt ctccaccgcg caggccccct ccaggtagaa ggccattttt ccaccctgta 5701 ggggaagccg attattttga ataccaccaa gaaggtggcc cagatggtga gcctgacgtg 5761 cccccgggag cgatagagca gggccccgca gatgacccag gagaaggccc aagcactgga 5821 ccccggggtc agggtgatgg aggcaggcgc aaaaaaggag ggtggtttgg aaagcatcgt 5881 ggtcaaggag gttccaaccc gaaatttgag aacattgcag aaggtttaag agctctcctg 5941 gctaggagtc acgtagaaag gactaccgac gaaggaactt gggtcgccgg tgtgttcgta 6001 tatggaggta gtaagacctc cctttacaac ctaaggcgag gaactgccct tgctattcca 6061 caatgtcgtc ttacaccatt gagtcgtctc ccctttggaa tggcccctgg acccggccca 6121 caacctggcc cgctaaggga gtccattgtc tgttatttca tggtcttttt acaaactcat 6181 atatttgctg aggttttgaa ggatgcgatt aaggaccttg ttatgacaaa gcccgctcct 6241 acctgcaata tcagggtgac tgtgtgcagc tttgacgatg gagtagattt gcctccctgg 6301 tttccaccta tggtggaagg ggctgccgcg gagggtgatg acggagatga cggagatgaa 6361 ggaggtgatg gagatgaggg tgaggaaggg caggagtgag gatccaaggg cg //