LOCUS       sequence-189190-        6263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Episomal reprogramming .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6263)
  AUTHORS   Howden SE, Maufort JP, Duffin BM, Elefanty AG, Stanley EG, Thomson 
            JA
  TITLE     Simultaneous Reprogramming and Gene Correction of Patient 
            Fibroblasts.
  JOURNAL   Stem Cell Reports. 2015 Dec 8;5(6):1109-18. doi: 
            10.1016/j.stemcr.2015.10.009. Epub 2015 Nov 12.
  PUBMED    26584543
REFERENCE   2  (bases 1 to 6263)
  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"; doi: 
            "10.1016/j.stemcr.2015.10.009"; journalName: "Stem Cell Reports"; 
            date: "2015-12-8- 8"; volume: "5"; issue: "6"; pages: "1109-18"
COMMENT     Sequence Label: pSimple-miR302/367
FEATURES             Location/Qualifiers
     source          1..6263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        213..1375
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          444..1366
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1323..1343
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(2553..2570)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2559..2783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2948..4061
                     /label=mini-oriP
                     /note="internally truncated variant of the Epstein-Barr 
                     virus oriP replication origin (Tanaka et al., 1999)"
     primer_bind     complement(4106..4122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4106..4122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4119..4141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4130..4146
                     /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(4154..4160,4161..4178,4179..4184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4199..4220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4337..4354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4508..5096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4588..4607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5267..6058,6059..6127))
                     /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     5890..5909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6128..6232)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg gaggctccgg tgcccgtcag tgggcagagc
      241 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
      301 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
      361 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
      421 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
      481 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc
      541 ttgatcccga gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc
      601 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
      661 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
      721 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat
      781 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
      841 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
      901 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
      961 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctccagg
     1021 gggctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     1081 gaaaggggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     1141 gtccaggcac ctcgattagt tctggagctt ttggagtacg tcgtctttag gttgggggga
     1201 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     1261 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     1321 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacg
     1381 tggtcgcggc caattctgca gtcgacggta ccgcgggccc gggatccgga attcccatcg
     1441 atggcctcga gaaagcggcc gcagctgcta gcgtccctgc cttttaccct tctggaggag
     1501 aacacgaatc tttgggaact agttcaggaa ggtaaaaaaa tttttttctt ctaaagttat
     1561 gccattttgt tttctttctc ctcagctcta aatactctga agtccaaaga agttgtatgt
     1621 tgggtgggct cccttcaact ttaacatgga agtgctttct gtgactttaa aagtaagtgc
     1681 ttccatgttt tagtaggagt gaatccaatt tacttctcca aaatagaaca cgctaacctc
     1741 atttgaaggg atcccctttg ctttaacatg ggggtacctg ctgtgtgaaa caaaagtaag
     1801 tgcttccatg tttcagtgga ggtgtctcca agccagcaca ccttttgtta caaaattttt
     1861 ttgttattgt gttttaaggt tactaagctt gttacaggtt aaaggattct aactttttcc
     1921 aagactgggc tccccaccac ttaaacgtgg atgtacttgc tttgaaacta aagaagtaag
     1981 tgcttccatg ttttggtgat ggtaagtctt ccttttacat ttttattatt tttttagaaa
     2041 ataactttat tgtattgacc gcagctcata tatttaagct ttattttgta tttttacatc
     2101 tgttaagggg ccccctctac tttaacatgg aggcacttgc tgtgacatga caaaaataag
     2161 tgcttccatg tttgagtgtg gtggttccta cctaatcagc aattgagtta acgcccacac
     2221 tgtgtgcagt tcttggctac aggccattac tgttgctaat atgcaactct gttgaatata
     2281 aattggaatt gcactttagc aatggtgatg gattgttaag ccaatgacag aatttaaacc
     2341 acagacttac tttgatagca ctcttaatgg tataacttct tctcccattt tatgtctctc
     2401 tttatgtttt ttcttatgtt tcctttttgt tttcaagaga gagctatctt ttagatctcc
     2461 agtatccttt tcctcttcag tagagatttc tatatcttga aataaataaa aaagctctcc
     2521 gttaagacat gggtctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     2581 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2641 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2701 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2761 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgaccga
     2821 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     2881 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccctggccgg
     2941 ggtcccgcag gaaaaggaca agcagcgaaa attcacgccc ccttgggagg tggcggcata
     3001 tgcaaaggat agcactccca ctctactact gggtatcata tgctgactgt atatgcatga
     3061 ggatagcata tgctacccgg atacagatta ggatagcata tactacccag atatagatta
     3121 ggatagcata tgctacccag atatagatta ggatagccta tgctacccag atataaatta
     3181 ggatagcata tactacccag atatagatta ggatagcata tgctacccag atatagatta
     3241 ggatagccta tgctacccag atatagatta ggatagcata tgctacccag atatagatta
     3301 ggatagcata tgctatccag atatttgggt agtatatgct acccagatat aaattaggat
     3361 agcatatact accctaatct ctattaggat agcatatgct acccggatac agattaggat
     3421 agcatatact acccagatat agattaggat agcatatgct acccagatat agattaggat
     3481 agcctatgct acccagatat aaattaggat agcatatact acccagatat agattaggat
     3541 agcatatgct acccagatat agattaggat agcctatgct acccagatat agattaggat
     3601 agcatatgct atccagatat ttgggtagta tatgctatcc atggcaacat tagcccaccg
     3661 tgctctcagc gacctcgtga atatgaggac caacaaccct gtgcttggcg ctcaggcgca
     3721 agtgtgtgta atttgtcctc cagatcgcag caatcgcgcc cctatcttgg cccgcccacc
     3781 tacttatgca ggtattcccc ggggtgccat tagtggtttt gtgggcaagt ggtttgaccg
     3841 cagtggttag cggggttaca atcagccaag ttattacacc cttattttac agtccaaaac
     3901 cgcagggcgg cgtgtggggg ctgacgcgat cgctgttcct taggaccctt ttactaaccc
     3961 taattcgata gcatatgctt cccgttgggt aacatatgct attgaattag ggttagtctg
     4021 gatagtatat actactaccc gggaagcata tgctacccgt ttagggttta ccgtcgacct
     4081 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4141 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4201 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4261 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4321 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5161 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5221 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5341 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5461 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5701 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5761 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5941 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6121 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6181 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6241 cccgaaaagt gccacctgac gtc
//
