LOCUS       sequence-155358-        6206 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 6206)
  AUTHORS   Aasen T, Raya A, Barrero MJ, Garreta E, Consiglio A, Gonzalez F, 
            Vassena R, Bilic J, Pekarik V, Tiscornia G, Edel M, Boue S, Belmonte
            JC
  TITLE     Efficient and rapid generation of induced pluripotent stem cells 
            from human keratinocytes.
  JOURNAL   Nat Biotechnol. 2008 Nov . 26(11):1276-84.
  PUBMED    18931654
REFERENCE   2  (bases 1 to 6206)
  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: "Nat 
            Biotechnol. 2008 Nov . 26(11):1276-84."
COMMENT     Sequence Label: pMSCV-Flag-hOct4
FEATURES             Location/Qualifiers
     source          1..6206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     365..387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     515..534
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             734..1250
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1314..1655
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1722..2138
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2068..2090
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2106..2122
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2154..2163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2163..2186
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             3334..3848
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4010..4026
                     /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(4034..4040,4041..4058,4059..4064))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4079..4100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4217..4234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5147..5938,5939..6007))
                     /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     5770..5789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6008..6112)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6180..6198
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
      241 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      301 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
      361 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
      421 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      481 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      541 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
      601 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
      661 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
      721 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
      781 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
      841 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
      901 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
      961 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     1021 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     1081 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     1141 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt
     1201 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
     1261 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
     1321 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     1381 tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     1441 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
     1501 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
     1561 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     1621 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
     1681 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     1741 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1801 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1861 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1921 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1981 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     2041 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     2101 cctcccttta tccagccctc actccttctc taggcgccgg aattagatct gccgccgcca
     2161 tggactacaa ggacgacgat gacaagctcg aggttaacga attcatggcg ggacacctgg
     2221 cttcggattt cgccttctcg ccccctccag gtggtggagg tgatgggcca ggggggccgg
     2281 agccgggctg ggttgatcct cggacctggc taagcttcca aggccctcct ggagggccag
     2341 gaatcgggcc gggggttggg ccaggctctg aggtgtgggg gattccccca tgccccccgc
     2401 cgtatgagtt ctgtgggggg atggcgtact gtgggcccca ggttggagtg gggctagtgc
     2461 cccaaggcgg cttggagacc tctcagcctg agggcgaagc aggagtcggg gtggagagca
     2521 actccgatgg ggcctccccg gagccctgca ccgtcacccc tggtgccgtg aagctggaga
     2581 aggagaagct ggagcaaaac ccggaggagt cccaggacat caaagctctg cagaaagaac
     2641 tcgagcaatt tgccaagctc ctgaagcaga agaggatcac cctgggatat acacaggccg
     2701 atgtggggct caccctgggg gttctatttg ggaaggtatt cagccaaacg accatctgcc
     2761 gctttgaggc tctgcagctt agcttcaaga acatgtgtaa gctgcggccc ttgctgcaga
     2821 agtgggtgga ggaagctgac aacaatgaaa accttcagga gatatgcaaa gcagaaaccc
     2881 tcgtgcaggc ccgaaagaga aagcgaacca gtatcgagaa ccgagtgaga ggcaacctgg
     2941 agaatttgtt cctgcagtgc ccgaaaccca cactgcagca gatcagccac atcgcccagc
     3001 agcttgggct cgagaaggat gtggtccgag tgtggttctg taaccggcgc cagaagggca
     3061 agcgatcaag cagcgactat gcacaacgag aggattttga ggctgctggg tctcctttct
     3121 cagggggacc agtgtccttt cctctggccc cagggcccca ttttggtacc ccaggctatg
     3181 ggagccctca cttcactgca ctgtactcct cggtcccttt ccctgagggg gaagcctttc
     3241 cccctgtctc tgtcaccact ctgggctctc ccatgcattc aaactgaatc gataaaataa
     3301 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     3361 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     3421 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     3481 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     3541 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     3601 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     3661 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     3721 agactgcgtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt
     3781 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     3841 tctttcatgg gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa
     3901 tattaagtaa tcctgactca attagccact gttttgaatc cacatactcc aatactcctg
     3961 aaatagttca ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc
     4021 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4081 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4141 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4201 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5281 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6121 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6181 ataggcgtat cacgaggccc tttcgt
//
