LOCUS       Exported                6536 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMSCV-Flag-hKlf4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  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 6536)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6536
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2163..2186
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             3664..4178
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4340..4356
                     /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(4364..4394)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4409..4430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4547..4564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4718..5306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4798..4817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5477..6337)
                     /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     6100..6119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6338..6442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6510..6528
                     /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 attcatggct gtcagcgacg
     2221 cgctgctccc atctttctcc acgttcgcgt ctggcccggc gggaagggag aagacactgc
     2281 gtcaagcagg tgccccgaat aaccgctggc gggaggagct ctcccacatg aagcgacttc
     2341 ccccagtgct tcccggccgc ccctatgacc tggcggcggc gaccgtggcc acagacctgg
     2401 agagcggcgg agccggtgcg gcttgcggcg gtagcaacct ggcgccccta cctcggagag
     2461 agaccgagga gttcaacgat ctcctggacc tggactttat tctctccaat tcgctgaccc
     2521 atcctccgga gtcagtggcc gccaccgtgt cctcgtcagc gtcagcctcc tcttcgtcgt
     2581 cgccgtcgag cagcggccct gccagcgcgc cctccacctg cagcttcacc tatccgatcc
     2641 gggccgggaa cgacccgggc gtggcgccgg gcggcacggg cggaggcctc ctctatggca
     2701 gggagtccgc tccccctccg acggctccct tcaacctggc ggacatcaac gacgtgagcc
     2761 cctcgggcgg cttcgtggcc gagctcctgc ggccagaatt ggacccggtg tacattccgc
     2821 cgcagcagcc gcagccgcca ggtggcgggc tgatgggcaa gttcgtgctg aaggcgtcgc
     2881 tgagcgcccc tggcagcgag tacggcagcc cgtcggtcat cagcgtcagc aaaggcagcc
     2941 ctgacggcag ccacccggtg gtggtggcgc cctacaacgg cgggccgccg cgcacgtgcc
     3001 ccaagatcaa gcaggaggcg gtctcttcgt gcacccactt gggcgctgga ccccctctca
     3061 gcaatggcca ccggccggct gcacacgact tccccctggg gcggcagctc cccagcagga
     3121 ctaccccgac cctgggtctt gaggaagtgc tgagcagcag ggactgtcac cctgccctgc
     3181 cgcttcctcc cggcttccat ccccacccgg ggcccaatta cccatccttc ctgcccgatc
     3241 agatgcagcc gcaagtcccg ccgctccatt accaagagct catgccaccc ggttcctgca
     3301 tgccagagga gcccaagcca aagaggggaa gacgatcgtg gccccggaaa aggaccgcca
     3361 cccacacttg tgattacgcg ggctgcggca aaacctacac aaagagttcc catctcaagg
     3421 cacacctgcg aacccacaca ggtgagaaac cttaccactg tgactgggac ggctgtggat
     3481 ggaaattcgc ccgctcagat gaactgacca ggcactaccg taaacacacg gggcaccgcc
     3541 cgttccagtg ccaaaaatgc gaccgagcat tttccaggtc ggaccacctc gccttacaca
     3601 tgaagaggca tttttaaatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     3661 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     3721 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     3781 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3841 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     3901 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3961 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     4021 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     4081 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     4141 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     4201 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     4261 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga
     4321 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     6481 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgt
//
