LOCUS       sequence-155853-        6077 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 6077)
  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 6077)
  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-hSox2
FEATURES             Location/Qualifiers
     source          1..6077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             72..588
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    652..993
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1060..1476
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1406..1428
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1444..1460
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1492..1501
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1501..1524
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             2543..3057
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3219..3235
                     /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(3243..3249,3250..3267,3268..3273))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3288..3309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3426..3443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3597..4185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3677..3696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4356..5147,5148..5216))
                     /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     4979..4998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5217..5321)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5389..5407
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5445..5467)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5567..5586
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5780..5802
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5930..5949
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag
       61 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
      121 ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa
      181 cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa
      301 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
      361 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
      421 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc
      481 cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg
      541 gagggtctcc tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac
      601 cgagatttgg agacccctgc ccagggacca ccgacccccc cgccgggagg taagctggcc
      661 agcggtcgtt tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg
      721 cgcctgcgtc tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
      781 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt
      841 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg
      901 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
      961 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt
     1021 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc
     1081 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1141 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1201 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1261 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     1321 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1381 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1441 tccctttatc cagccctcac tccttctcta ggcgccggaa ttagatctgc cgccgccatg
     1501 gactacaagg acgacgatga caagctcgag gttaacgaat tcatgtacaa catgatggag
     1561 acggagctga agccgccggg cccgcagcaa acttcggggg gcggcggcgg caactccacc
     1621 gcggcggcgg ccggcggcaa ccagaaaaac agcccggacc gcgtcaagcg gcccatgaat
     1681 gccttcatgg tgtggtcccg cgggcagcgg cgcaagatgg cccaggagaa ccccaagatg
     1741 cacaactcgg agatcagcaa gcgcctgggc gccgagtgga aacttttgtc ggagacggag
     1801 aagcggccgt tcatcgacga ggctaagcgg ctgcgagcgc tgcacatgaa ggagcacccg
     1861 gattataaat accggccccg gcggaaaacc aagacgctca tgaagaagga taagtacacg
     1921 ctgcccggcg ggctgctggc ccccggcggc aatagcatgg cgagcggggt cggggtgggc
     1981 gccggcctgg gcgcgggcgt gaaccagcgc atggacagtt acgcgcacat gaacggctgg
     2041 agcaacggca gctacagcat gatgcaggac cagctgggct acccgcagca cccgggcctc
     2101 aatgcgcacg gcgcagcgca gatgcagccc atgcaccgct acgacgtgag cgccctgcag
     2161 tacaactcca tgaccagctc gcagacctac atgaacggct cgcccaccta cagcatgtcc
     2221 tactcgcagc agggcacccc tggcatggct cttggctcca tgggttcggt ggtcaagtcc
     2281 gaggccagct ccagcccccc tgtggttacc tcttcctccc actccagggc gccctgccag
     2341 gccggggacc tccgggacat gatcagcatg tatctccccg gcgccgaggt gccggaaccc
     2401 gccgccccca gcagacttca catgtcccag cactaccaga gcggcccggt gcccggcacg
     2461 gccattaacg gcacactgcc cctctcacac atgtgaatcg ataaaataaa agattttatt
     2521 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     2581 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
     2641 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     2701 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     2761 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     2821 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     2881 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
     2941 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     3001 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg
     3061 taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat
     3121 cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat
     3181 tatggacagc gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca
     3241 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3301 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3361 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3421 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3481 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3541 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3601 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3661 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3721 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3781 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3841 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3901 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3961 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4021 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4081 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4141 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4201 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4261 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4321 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4381 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4441 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4501 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4561 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4621 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4681 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4741 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4801 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4861 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4921 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4981 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5041 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5101 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5161 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5221 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5281 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5341 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5401 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5461 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5521 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     5581 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5641 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5701 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     5761 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca aggaatggtg
     5821 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     5881 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     5941 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6001 cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag gctctagttt
     6061 tgactcaaca atatcac
//
