LOCUS       Exported                7633 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMSCVpuro-Flag-cMyc T58A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7633)
  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 7633)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7633
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(941..1801)
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1974..1992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2030..2052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2152..2171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2365..2387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2515..2534
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2734..3250
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3314..3655
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3722..4138
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4068..4090
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4106..4122
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      4154..4163
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4163..4186
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4193..5509
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="PLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAPS
                     EDIWKKFELLPAPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTEL
                     LGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPNP
                     ARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDSL
                     LSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGSP
                     SAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNRK
                     CTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKAT
                     AYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     promoter        5542..6041
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5577..5600)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5974..5993
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6062..6661
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6062..6081)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6558..6578
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6761..7275
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7437..7453
                     /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(7461..7491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7506..7527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa
     2401 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     2461 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     2521 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     2581 tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     2641 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     2701 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     2761 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     2821 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     2881 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     2941 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3001 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3061 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     3121 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa
     3181 tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg
     3241 gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc
     3301 cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt
     3361 ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat
     3421 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
     3481 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
     3541 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     3601 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     3661 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     3721 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     3781 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     3841 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     3901 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     3961 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     4021 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     4081 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg
     4141 aattagatct gccgccgcca tggactacaa ggacgacgat gacaagctcg agcccctcaa
     4201 cgttagcttc accaacagga actatgacct cgactacgac tcggtgcagc cgtatttcta
     4261 ctgcgacgag gaggagaact tctaccagca gcagcagcag agcgagctgc agcccccggc
     4321 gcccagcgag gatatctgga agaaattcga gctgctgccc gcgccgcccc tgtcccctag
     4381 ccgccgctcc gggctctgct cgccctccta cgttgcggtc acacccttct cccttcgggg
     4441 agacaacgac ggcggtggcg ggagcttctc cacggccgac cagctggaga tggtgaccga
     4501 gctgctggga ggagacatgg tgaaccagag tttcatctgc gacccggacg acgagacctt
     4561 catcaaaaac atcatcatcc aggactgtat gtggagcggc ttctcggccg ccgccaagct
     4621 cgtctcagag aagctggcct cctaccaggc tgcgcgcaaa gacagcggca gcccgaaccc
     4681 cgcccgcggc cacagcgtct gctccacctc cagcttgtac ctgcaggatc tgagcgccgc
     4741 cgcctcagag tgcatcgacc cctcggtggt cttcccctac cctctcaacg acagcagctc
     4801 gcccaagtcc tgcgcctcgc aagactccag cgccttctct ccgtcctcgg attctctgct
     4861 ctcctcgacg gagtcctccc cgcagggcag ccccgagccc ctggtgctcc atgaggagac
     4921 accgcccacc accagcagcg actctgagga ggaacaagaa gatgaggaag aaatcgatgt
     4981 tgtttctgtg gaaaagaggc aggctcctgg caaaaggtca gagtctggat caccttctgc
     5041 tggaggccac agcaaacctc ctcacagccc actggtcctc aagaggtgcc acgtctccac
     5101 acatcagcac aactacgcag cgcctccctc cactcggaag gactatcctg ctgccaagag
     5161 ggtcaagttg gacagtgtca gagtcctgag acagatcagc aacaaccgaa aatgcaccag
     5221 ccccaggtcc tcggacaccg aggagaatgt caagaggcga acacacaacg tcttggagcg
     5281 ccagaggagg aacgagctaa aacggagctt ttttgccctg cgtgaccaga tcccggagtt
     5341 ggaaaacaat gaaaaggccc ccaaggtagt tatccttaaa aaagccacag catacatcct
     5401 gtccgtccaa gcagaggagc aaaagctcat ttctgaagag gacttgttgc ggaaacgacg
     5461 agaacagttg aaacacaaac ttgaacagct acggaactct tgtgcgtaaa tcgatgcata
     5521 aaatcactag tgaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     5581 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     5641 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     5701 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     5761 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     5821 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     5881 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     5941 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     6001 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac
     6061 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     6121 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     6181 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     6241 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     6301 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     6361 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     6421 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     6481 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     6541 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     6601 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     6661 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     6721 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     6781 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     6841 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     6901 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     6961 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     7021 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     7081 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     7141 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     7201 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     7261 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     7321 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     7381 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     7441 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7501 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7561 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7621 cgtattgggc gct
//
