LOCUS       40924_14060        7037 bp DNA     circular SYN 07-AUG-2018
DEFINITION  Non-replicating episomal expression of EBNA1, which enhances 
            transfection efficiency and expression from episomal plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7037)
  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 7037)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7037)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells.
            2012 Nov 29. doi: 10.1002/stem.1293."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids,
                     forward primer"
     CDS             1747..3669
                     /codon_start=1
                     /label=EBNA1
                     /note="Epstein-Barr nuclear antigen 1, also known as
                     EBNA-1"
                     /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
                     GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
                     AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA
                     GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
                     GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG
                     GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     misc_feature    3716..4304
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(4394..4413)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4459..4514
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4513..4532)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also
                     called rb-glob-pA-term-R"
     primer_bind     complement(4875..4891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4899..4915)
                     /label=lac operator
                     /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(4923..4953)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4968..4989)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(5114..5131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5285..5873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6047..6904)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6905..7009)
                     /label=AmpR promoter
     promoter        7037
                     /label=CAG
                     /note="CMV early enhancer fused to modified chicken 
                     beta-actin promoter"
ORIGIN
        1 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
      421 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
      481 gatgggggcg gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcgggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgcccttgtc
     1741 gacaccatgt ctgacgaggg gccaggtaca ggacctggaa atggcctagg agagaaggga
     1801 gacacatctg gaccagaagg ctccggcggc agtggacctc aaagaagagg gggtgataac
     1861 catggacgag gacggggaag aggacgagga cgaggaggcg gaagaccagg agccccgggc
     1921 ggctcaggat cagggccaag acatagagat ggtgtccgga gaccccaaaa acgtccaagt
     1981 tgcattggct gcaaagggac ccacggtgga acaggagcag gagcaggagc gggaggggca
     2041 ggagcaggag gggcaggagc aggaggaggg gcaggagcag gaggaggggc aggaggggca
     2101 ggaggggcag gaggggcagg agcaggagga ggggcaggag caggaggagg ggcaggaggg
     2161 gcaggagggg caggagcagg aggaggggca ggagcaggag gaggggcagg aggggcagga
     2221 gcaggaggag gggcaggagg ggcaggaggg gcaggagcag gaggaggggc aggagcagga
     2281 ggaggggcag gaggggcagg agcaggagga ggggcaggag gggcaggagg ggcaggagca
     2341 ggaggagggg caggagcagg aggggcagga ggggcaggag gggcaggagc aggaggggca
     2401 ggagcaggag gaggggcagg aggggcagga ggggcaggag caggaggggc aggagcagga
     2461 ggggcaggag caggaggggc aggagcagga ggggcaggag gggcaggagc aggaggggca
     2521 ggaggggcag gagcaggagg ggcaggaggg gcaggagcag gaggaggggc aggaggggca
     2581 ggagcaggag gaggggcagg aggggcagga gcaggagggg caggaggggc aggagcagga
     2641 ggggcaggag gggcaggagc aggaggggca ggaggggcag gagcaggagg aggggcagga
     2701 gcaggagggg caggagcagg aggtggaggc cggggtcgag gaggcagtgg aggccggggt
     2761 cgaggaggta gtggaggccg gggtcgagga ggtagtggag gccgccgggg tagaggacgt
     2821 gaaagagcca gggggggaag tcgtgaaaga gccaggggga gaggtcgtgg acgtggagaa
     2881 aagaggccca ggagtcccag tagtcagtca tcatcatccg ggtctccacc gcgcaggccc
     2941 cctccaggta gaaggccatt tttccaccct gtaggggaag ccgattattt tgaataccac
     3001 caagaaggtg gcccagatgg tgagcctgac gtgcccccgg gagcgataga gcagggcccc
     3061 gcagatgacc caggagaagg cccaagcact ggaccccggg gtcagggtga tggaggcagg
     3121 cgcaaaaaag gagggtggtt tggaaagcat cgtggtcaag gaggttccaa cccgaaattt
     3181 gagaacattg cagaaggttt aagagctctc ctggctagga gtcacgtaga aaggactacc
     3241 gacgaaggaa cttgggtcgc cggtgtgttc gtatatggag gtagtaagac ctccctttac
     3301 aacctaaggc gaggaactgc ccttgctatt ccacaatgtc gtcttacacc attgagtcgt
     3361 ctcccctttg gaatggcccc tggacccggc ccacaacctg gcccgctaag ggagtccatt
     3421 gtctgttatt tcatggtctt tttacaaact catatatttg ctgaggtttt gaaggatgcg
     3481 attaaggacc ttgttatgac aaagcccgct cctacctgca atatcagggt gactgtgtgc
     3541 agctttgacg atggagtaga tttgcctccc tggtttccac ctatggtgga aggggctgcc
     3601 gcggagggtg atgacggaga tgacggagat gaaggaggtg atggagatga gggtgaggaa
     3661 gggcaggagt gaggatccaa gggcgaattc aagcttcggg gactagtcat atgataatca
     3721 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3781 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3841 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3901 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3961 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4021 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4081 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     4141 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4201 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4261 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcggt aaattcactc
     4321 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     4381 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     4441 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4501 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     4561 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     4621 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4681 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4741 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     4801 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc
     4861 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4921 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4981 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5041 cggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7021 gaaaagtgcc acctggg
//
