LOCUS       sequence-354143-        8496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-integrating (episomal) expression of mouse Oct3/4, Klf4 and 
            Sox2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8496)
  AUTHORS   Okita K, Nakagawa M, Hyenjong H, Ichisaka T, Yamanaka S
  TITLE     Generation of mouse induced pluripotent stem cells without viral 
            vectors.
  JOURNAL   Science. 2008 Nov 7. 322(5903):949-53.
  PUBMED    18845712
REFERENCE   2  (bases 1 to 8496)
  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: "Science. 
            2008 Nov 7. 322(5903):949-53."
COMMENT     Sequence Label: pCX-OKS-2A
FEATURES             Location/Qualifiers
     source          1..8496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2843..2908
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             4358..4423
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     primer_bind     complement(5517..5536)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5582..5637
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5636..5655)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5998..6014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5998..6014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6011..6033)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6022..6038
                     /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(6046..6052,6053..6070,6071..6076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6091..6112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6171..6366
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6217..6352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6372..6506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6409..6428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6463..6482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6574..6591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6745..7333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6825..6844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7504..8295,8296..8364))
                     /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     8127..8146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8365..8469)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggs gggggggggg ggggggsgsg cscsrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs grggsggrgr ggkgsggsgg srgccaawca grgsggsgcg cyccgaaagt
      601 ttccttttwt ggcgrggsgg sggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggkggcggcr ggkgggggkg ccgggsgggg
     1261 sggggccgcc ycgggccggg grgggckcgg gggrggggcg cggcggcccc cggrgcgccg
     1321 gcggctgtcg rggcgcggcg rgccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcgccctta
     1741 aaaagcaggc tccaccttcc ccatggctgg acacctggct tcagacttcg ccttctcacc
     1801 cccaccaggt gggggtgatg ggtcagcagg gctggagccg ggctgggtgg attctcgaac
     1861 ctggctaagc ttccaagggc ctccaggtgg gcctggaatc ggaccaggct cagaggtatt
     1921 ggggatctcc ccatgtccgc ccgcatacga gttctgcgga gggatggcat actgtggacc
     1981 tcaggttgga ctgggcctag tcccccaagt tggcgtggag actttgcagc ctgagggcca
     2041 ggcaggagca cgagtggaaa gcaactcaga gggaacctcc tctgagccct gtgccgaccg
     2101 ccccaatgcc gtgaagttgg agaaggtgga accaactccc gaggagtccc aggacatgaa
     2161 agccctgcag aaggagctag aacagtttgc caagctgctg aagcagaaga ggatcacctt
     2221 ggggtacacc caggccgacg tggggctcac cctgggcgtt ctctttggaa aggtgttcag
     2281 ccagaccacc atctgtcgct tcgaggcctt gcagctcagc cttaagaaca tgtgtaagct
     2341 gcggcccctg ctggagaagt gggtggagga agccgacaac aatgagaacc ttcaggagat
     2401 atgcaaatcg gagaccctgg tgcaggcccg gaagagaaag cgaactagca ttgagaaccg
     2461 tgtgaggtgg agtctggaga ccatgtttct gaagtgcccg aagccctccc tacagcagat
     2521 cactcacatc gccaatcagc ttgggctaga gaaggatgtg gttcgagtat ggttctgtaa
     2581 ccggcgccag aagggcaaaa gatcaagtat tgagtattcc caacgagaag agtatgaggc
     2641 tacagggaca cctttcccag ggggggctgt atcctttcct ctgcccccag gtccccactt
     2701 tggcacccca ggctatggaa gcccccactt caccacactc tactcagtcc cttttcctga
     2761 gggcgaggcc tttccctctg ttcccgtcac tgctctgggc tctcccatgc attcaaacgg
     2821 ggatctcaaa attgtcgctc ctgtcaaaca aactcttaac tttgatttac tcaaactggc
     2881 tggggatgta gaaagcaatc caggtccact catggctgtc agcgacgctc tgctcccgtc
     2941 cttctccacg ttcgcgtccg gcccggcggg aagggagaag acactgcgtc cagcaggtgc
     3001 cccgactaac cgttggcgtg aggaactctc tcacatgaag cgacttcccc cacttcccgg
     3061 ccgcccctac gacctggcgg cgacggtggc cacagacctg gagagtggcg gagctggtgc
     3121 agcttgcagc agtaacaacc cggccctcct agcccggagg gagaccgagg agttcaacga
     3181 cctcctggac ctagacttta tcctttccaa ctcgctaacc caccaggaat cggtggccgc
     3241 caccgtgacc acctcggcgt cagcttcatc ctcgtcttcc ccagcgagca gcggccctgc
     3301 cagcgcgccc tccacctgca gcttcagcta tccgatccgg gccgggggtg acccgggcgt
     3361 ggctgccagc aacacaggtg gagggctcct ctacagccga gaatctgcgc cacctcccac
     3421 ggcccccttc aacctggcgg acatcaatga cgtgagcccc tcgggcggct tcgtggctga
     3481 gctcctgcgg ccggagttgg acccagtata cattccgcca cagcagcctc agccgccagg
     3541 tggcgggctg atgggcaagt ttgtgctgaa ggcgtctctg accacccctg gcagcgagta
     3601 cagcagccct tcggtcatca gtgttagcaa aggaagccca gacggcagcc accccgtggt
     3661 agtggcgccc tacagcggtg gcccgccgcg catgtgcccc aagattaagc aagaggcggt
     3721 cccgtcctgc acggtcagcc ggtccctaga ggcccatttg agcgctggac cccagctcag
     3781 caacggccac cggcccaaca cacacgactt ccccctgggg cggcagctcc ccaccaggac
     3841 tacccctaca ctgagtcccg aggaactgct gaacagcagg gactgtcacc ctggcctgcc
     3901 tcttccccca ggattccatc cccatccggg gcccaactac cctcctttcc tgccagacca
     3961 gatgcagtca caagtcccct ctctccatta tcaagagctc atgccaccgg gttcctgcct
     4021 gccagaggag cccaagccaa agaggggaag aaggtcgtgg ccccggaaaa gaacagccac
     4081 ccacacttgt gactatgcag gctgtggcaa aacctatacc aagagttctc atctcaaggc
     4141 acacctgcga actcacacag gcgagaaacc ttaccactgt gactgggacg gctgtgggtg
     4201 gaaattcgcc cgctccgatg aactgaccag gcactaccgc aaacacacag ggcaccggcc
     4261 ctttcagtgc cagaagtgcg acagggcctt ttccaggtcg gaccaccttg ccttacacat
     4321 gaagaggcac tttggggatc tcaaaattgt cgctcctgtc aaacaaactc ttaactttga
     4381 tttactcaaa ctggctgggg atgtagaaag caatccaggt ccactcatga acaattcgcc
     4441 cttcaccatg tataacatga tggagacgga gctgaagccg ccgggcccgc agcaagcttc
     4501 ggggggcggc ggcggaggag gcaacgccac ggcggcggcg accggcggca accagaagaa
     4561 cagcccggac cgcgtcaaga ggcccatgaa cgccttcatg gtatggtccc gggggcagcg
     4621 gcgtaagatg gcccaggaga accccaagat gcacaactcg gagatcagca agcgcctggg
     4681 cgcggagtgg aaacttttgt ccgagaccga gaagcggccg ttcatcgacg aggccaagcg
     4741 gctgcgcgct ctgcacatga aggagcaccc ggattataaa taccggccgc ggcggaaaac
     4801 caagacgctc atgaagaagg ataagtacac gcttcccgga ggcttgctgg cccccggcgg
     4861 gaacagcatg gcgagcgggg ttggggtggg cgccggcctg ggtgcgggcg tgaaccagcg
     4921 catggacagc tacgcgcaca tgaacggctg gagcaacggc agctacagca tgatgcagga
     4981 gcagctgggc tacccgcagc acccgggcct caacgctcac ggcgcggcac agatgcaacc
     5041 gatgcaccgc tacgacgtca gcgccctgca gtacaactcc atgaccagct cgcagaccta
     5101 catgaacggc tcgcccacct acagcatgtc ctactcgcag cagggcaccc ccggtatggc
     5161 gctgggctcc atgggctctg tggtcaagtc cgaggccagc tccagccccc ccgtggttac
     5221 ctcttcctcc cactccaggg cgccctgcca ggccggggac ctccgggaca tgatcagcat
     5281 gtacctcccc ggcgccgagg tgccggagcc cgctgcgccc agtagactgc acatggccca
     5341 gcactaccag agcggcccgg tgcccggcac ggccattaac ggcacactgc ccctgtcgca
     5401 catgtgaggg ctggaagggc gaattgcctg caggaattca ctcctcaggt gcaggctgcc
     5461 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     5521 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     5581 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     5641 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     5701 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     5761 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     5821 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     5881 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     5941 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     6001 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6061 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6121 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     6181 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6241 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6301 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6361 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     6421 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6481 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     6541 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6601 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6661 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6721 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6781 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6841 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6901 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6961 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7021 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7081 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7141 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7201 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7261 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7321 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7381 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7441 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7501 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7561 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7621 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7681 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7741 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7801 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7861 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7921 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7981 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8041 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8101 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8161 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8221 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8281 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8341 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8401 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8461 ggttccgcgc acatttcccc gaaaagtgcc acctgg
//
