LOCUS       sequence-340919-        6412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells.
            2012 Nov 29. doi: 10.1002/stem.1293."
COMMENT     Sequence Label: pCXB-EBNA1
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(612..618,619..636,637..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(join(1733..2524,2525..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
//
