LOCUS       sequence-148223-        9879 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A modified episomal (EBNA1/oriP) vector expressing human OCT4 and 
            SOX2 genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9879)
  AUTHORS   Chou BK, Gu H, Gao Y, Dowey SN, Wang Y, Shi J, Li Y, Ye Z, Cheng T, 
            Cheng L
  TITLE     A Facile Method to Establish Human Induced Pluripotent Stem Cells 
            From Adult Blood Cells Under Feeder-Free and Xeno-Free Culture 
            Conditions: A Clinically Compliant Approach.
  JOURNAL   Stem Cells Transl Med. 2015 Mar 5. pii: sctm.2014-0214.
  PUBMED    25742692
REFERENCE   2  (bases 1 to 9879)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells 
            Transl Med. 2015 Mar 5. pii: sctm.2014-0214."
COMMENT     Sequence Label: MOS
FEATURES             Location/Qualifiers
     source          1..9879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        120..527
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             1621..1680
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             2650..2679
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2698..2721
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2791..2811)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3209..3220)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3317..3451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3341..3360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3395..3414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3594..3616)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      4179..5968
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(8006..8024)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8092..8196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8197..8265,8266..9057)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8415..8434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9228..9816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9717..9736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
       61 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgag ctagctgcag
      121 taacgccatt ttgcaaggca tggaaaaata ccaaaccaag aatagagaag ttcagatcaa
      181 gggcgggtac atgaaaatag ctaacgttgg gccaaacagg atatctgcgg tgagcagttt
      241 cggccccggc ccggggccaa gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc
      301 aagaacagat ggtccccaga tatggcccaa ccctcagcag tttcttaaga cccatcagat
      361 gtttccaggc tcccccaagg acctgaaatg accctgcgcc ttatttgaat taaccaatca
      421 gcctgcttct cgcttctgtt cgcgcgcttc tgcttcccga gctctataaa agagctcaca
      481 acccctcact cggcgcgcca gtcctccgac agactgagtc gcccgggtac cgcgatcgcc
      541 atggcgggac acctggcttc ggatttcgcc ttctcgcccc ctccaggtgg tggaggtgat
      601 gggccagggg ggccggagcc gggctgggtt gatcctcgga cctggctaag cttccaaggc
      661 cctcctggag ggccaggaat cgggccgggg gttgggccag gctctgaggt gtgggggatt
      721 cccccatgcc ccccgccgta tgagttctgt ggggggatgg cgtactgtgg gccccaggtt
      781 ggagtggggc tagtgcccca aggcggcttg gagacctctc agcctgaggg tgaagcagga
      841 gtcggggtgg agagcaactc cgatggggcc tccccggagc cctgcaccgt cacccctggt
      901 gccgtgaagc tggagaagga gaagctggag caaaacccgg aggagtccca ggacatcaaa
      961 gctctgcaga aagaactcga gcaatttgcc aagctcctga agcagaagag gatcaccctg
     1021 ggatatacac aggccgatgt ggggctcacc ctgggggttc tatttgggaa ggtattcagc
     1081 caaacgacca tctgccgctt tgaggctctg cagcttagct tcaagaacat gtgtaagctg
     1141 cggcccttgc tgcagaagtg ggtggaggaa gctgacaaca atgaaaatct tcaggagata
     1201 tgcaaagcag aaaccctcgt gcaggcccga aagagaaagc gaaccagtat cgagaaccga
     1261 gtgagaggca acctggagaa tttgttcctg cagtgcccga aacccacact gcagcagatc
     1321 agccacatcg cccagcagct tgggctcgag aaggatgtgg tccgagtgtg gttctgtaac
     1381 cggcgccaga agggcaagcg atcaagcagc gactatgcac aacgagagga ttttgaggct
     1441 gctgggtctc ctttctcagg gggaccagtg tcctttcctc tggccccagg gccccatttt
     1501 ggtaccccag gctatgggag ccctcacttc actgcactgt actcctcggt ccctttccct
     1561 gagggggaag cctttccccc tgtctccgtc accactctgg gctctcccat gcattcaaac
     1621 cagtgtacta attatgctct cttgaaattg gctggagatg ttgagagcaa cccaggtccc
     1681 atgtacaaca tgatggagac ggagctgaag ccgccgggcc cgcagcaaac ttcggggggc
     1741 ggcggcggca actccaccgc ggcggcggcc ggcggcaacc agaaaaacag cccggaccgc
     1801 gtcaagcggc ccatgaatgc cttcatggtg tggtcccgcg ggcagcggcg caagatggcc
     1861 caggagaacc ccaagatgca caactcggag atcagcaagc gcctgggcgc cgagtggaaa
     1921 cttttgtcgg agacggagaa gcggccgttc atcgacgagg ctaagcggct gcgagcgctg
     1981 cacatgaagg agcacccgga ttataaatac cggccccggc ggaaaaccaa gacgctcatg
     2041 aagaaggata agtacacgct gcccggcggg ctgctggccc ccggcggcaa tagcatggcg
     2101 agcggggtcg gggtgggcgc cggcctgggc gcgggcgtga accagcgcat ggacagttac
     2161 gcgcacatga acggctggag caacggcagc tacagcatga tgcaggacca gctgggctac
     2221 ccgcagcacc cgggcctcaa tgcgcacggc gcagcgcaga tgcagcccat gcaccgctac
     2281 gacgtgagcg ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg
     2341 cccacctaca gcatgtccta ctcgcagcag ggcacccctg gcatggctct tggctccatg
     2401 ggttcggtgg tcaagtccga ggccagctcc agcccccctg tggttacctc ttcctcccac
     2461 tccagggcgc cctgccaggc cggggacctc cgggacatga tcagcatgta tctccccggc
     2521 gccgaggtgc cggaacccgc cgcccccagc agacttcaca tgtcccagca ctaccagagc
     2581 ggcccggtgc ccggcacggc cattaacggc acactgcccc tctcacacat gacgcgtacg
     2641 cggccgctcg agcagaaact catctcagaa gaggatctgg cagcaaatga tatcctggat
     2701 tacaaggatg acgacgataa ggtttaaacg cgtcgacaat caacctctgg attacaaaat
     2761 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2821 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2881 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2941 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3001 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3061 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3121 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     3181 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3241 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3301 gatctccctt tgggcggatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     3361 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3421 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3481 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3541 tatggctgat tatgatccgg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     3601 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     3661 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gaggtcgact
     3721 ctactagagg atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc
     3781 ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc
     3841 cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg
     3901 acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag
     3961 actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc
     4021 tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct
     4081 acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca
     4141 ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg
     4201 ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga
     4261 agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac
     4321 cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca
     4381 ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct
     4441 tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag
     4501 taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt
     4561 ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct
     4621 tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc
     4681 atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg
     4741 gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg
     4801 accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac
     4861 gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc
     4921 acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt
     4981 gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt
     5041 aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac
     5101 cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc
     5161 gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt
     5221 tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta
     5281 gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat
     5341 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     5401 atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     5461 atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat
     5521 atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat
     5581 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc
     5641 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     5701 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     5761 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     5821 tatcctaatc tgtatccggg tagcatatgc tatcctcatg catatacagt cagcatatga
     5881 tacccagtag tagagtggga gtgctatcct ttgcatatgc cgccacctcc caagggggcg
     5941 tgaattttcg ctgcttgtcc ttttcctgct ggttgctccc attcttaggt gaatttaagg
     6001 aggccaggct aaagccgtcg catgtctgat tgctcaccag gtaaatgtcg ctaatgtttt
     6061 ccaacgcgag aaggtgttga gcgcggagct gagtgacgtg acaacatggg tatgcccaat
     6121 tgccccatgt tgggaggacg aaaatggtga caagacagat ggccagaaat acaccaacag
     6181 cacgcatgat gtctactggg gatttattct ttagtgcggg ggaatacacg gcttttaata
     6241 cgattgaggg cgtctcctaa caagttacat cactcctgcc cttcctcacc ctcatctcca
     6301 tcacctcctt catctccgtc atctccgtca tcaccctccg cggcagcccc ttccaccata
     6361 ggtggaaacc agggaggcaa atctactcca tcgtcaaagc tgcacacagt caccctgata
     6421 ttgcaggtag gagcgggctt tgtcataaca aggtccttaa tcgcatcctt caaaacctca
     6481 gcaaatatat gagtttgtaa aaagaccatg aaataacaga caatggactc ccttagcggg
     6541 ccaggttgtg ggccgggtcc aggggccatt ccaaagggga gacgactcaa tggtgtaaga
     6601 cgacattgtg gaatagcaag ggcagttcct cgccttaggt tgtaaaggga ggtcttacta
     6661 cctccatata cgaacacacc ggcgacccaa gttccttcgt cggtagtcct ttctacgtga
     6721 ctcctagcca ggagagctct taaaccttct gcaatgttct caaatttcgg gttggaacct
     6781 ccttgaccac gatgctttcc aaaccaccct ccttttttgc gcctgcctcc atcaccctga
     6841 ccccggggtc cagtgcttgg gccttctcct gggtcatctg cggggccctg ctctatcgct
     6901 cccgggggca cgtcaggctc accatctggg ccaccttctt ggtggtattc aaaataatcg
     6961 gcttccccta cagggtggaa aaatggcctt ctacctggag ggggcctgcg cggtggagac
     7021 ccggatgatg atgactgact actgggactc ctgggcctct tttctccacg tccacgacct
     7081 ctccccctgg ctctttcacg acttcccccc ctggctcttt cacgtcctct accccggcgg
     7141 cctccactac ctcctcgacc ccggcctcca ctacctcctc gaccccggcc tccactgcct
     7201 cctcgacccc ggcctccacc tcctgctcct gcccctcctg ctcctgcccc tcctcctgct
     7261 cctgcccctc ctgctcctgc ccctcccgct cctgctcctg ctcctgttcc accgtgggtc
     7321 cctttgcagc caatgcaact tggacgtttt tggggtctcc ggacaccatc tctatgtctt
     7381 ggccctgatc ctgagccgcc cggggctcct ggtcttccgc ctcctcgtcc tcgtcctctt
     7441 ccccgtcctc gtccatggtt atcaccccct cttctttgag gtccactgcc gccggagcct
     7501 tctggtccag atgtgtctcc cttctctcct aggccatttc caggtcctgt acctggcccc
     7561 tcgtcagaca tgattcacac taaaagagat caatagacat ctttattaga cgacgctcag
     7621 tgaatacagg gagtgcagac tcctgccccc tccaacagcc cccccaccct catccccttc
     7681 atggtcgctg tcagacagat ccaggtctga aaattcccca tcctccgaac catcctcgtc
     7741 ctcatcacca attactcgca gcccggaaaa ctcccgctga acatcctcaa gatttgcgtc
     7801 ctgagcctca agccaggcct caaattcctc gtcccccttt ttgctggacg gtagggatgg
     7861 ggattctcgg gacccctcct cttcctcttc aaggtcacca gacagagatg ctactggggc
     7921 aacggaagaa aagctgggtg cggcctgtga ggatcagctt atcgatgata agctgtcaaa
     7981 catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     8041 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     8101 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     8161 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     8221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     8281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     8341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     8401 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     8461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     8521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     8581 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     8641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     8701 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     8761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     8821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     8881 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     8941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     9001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     9061 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     9121 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     9181 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     9241 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     9301 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     9361 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     9421 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     9481 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     9541 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     9601 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     9661 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     9721 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     9781 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     9841 cggttcctgg ccttttgctg gccttgaagc tgtccctga
//
