LOCUS       sequence-13884-e        5621 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 5621)
  AUTHORS   Chen G, Gulbranson DR, Hou Z, Bolin JM, Ruotti V, Probasco MD, 
            Smuga-Otto K, Howden SE, Diol NR, Propson NE, Wagner R, Lee GO, 
            Antosiewicz-Bourget J, Teng JM, Thomson JA
  TITLE     Chemically defined conditions for human iPSC derivation and culture.
  JOURNAL   Nat Methods. 2011 May;8(5):424-9. doi: 10.1038/nmeth.1593. Epub 2011
            Apr 10.
  PUBMED    21478862
REFERENCE   2  (bases 1 to 5621)
  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: 
            "10.1038/nmeth.1593"; journalName: "Nat Methods"; date: "2011-05"; 
            volume: "8"; issue: "5"; pages: "424-9"
COMMENT     Sequence Label: pET-3c-rhVTN-NC
FEATURES             Location/Qualifiers
     source          1..5621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(404..451)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     447..465
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             1540..1562
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1593..1612)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1594..1612)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1696..1715)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3173..3364
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3358..3380)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3466..3608
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3521..3540
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3623..3640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3794..4382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3874..3893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4553..5344,5345..5413))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5176..5195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5414..5518)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5586..5604
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
      241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
      301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
      361 cacacccgtc ctgtggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
      421 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact
      481 cagcttcctt tcgggctttg ttagcagccg gatccctagc gggatggccg gcgggagttc
      541 tggttgcggc cacggctgtg gcctcgttgt gaacggtagc ctttgcggtt gcgatgccta
      601 aacctttgtt tcttggccaa ggaggggcgg ggtgccatgc ctgagatgta gatgcggcca
      661 gccatggctg cgtccacttg ccctggcaca ccgtgccagt cccggctaat gaactggggc
      721 tgtctggtac cagcagaggt tctgccccag aagagaagct cgaagatgtc ctcccagctg
      781 tcccgctgca tcatggcaaa gtgttcaaac acagccgaca gggagctgcc ttcacactcc
      841 tcctgactgg gctggtgctg gaactggtac tcccagtact gtttcccctt gaagaagtag
      901 acccgctccc ggccactgta gctatgggca gggagggcca aggctgcatc cacgttgtcc
      961 gggatgccat cgaagccgtc agagatattt cgggggtaat cagggtccag gacaccatcc
     1021 tcaaagcgcc agtactgact acccttgaag aggtaggtct tcccctgaca gttgatgcgg
     1081 gtgaaggcgg catcgatggg gccctcgatg ccccagacat ctcggatgag cttggggtac
     1141 ccaggcctca ctgccttttc gtccagttca tagcagtact gccctcggaa ggcaaagagg
     1201 gaaccgttct tgaggtcggt gaaggcgtcg aagggcttcc cactgcacag ctcctcctct
     1261 gctgggggct gaggtctccc tggatgaagg gtctcaggcc ttgagtctat cccctcaggc
     1321 ttagaggcgc ccacctcagg cgcaggggcc tcttcctcag gtttcagaac aggtgtctgc
     1381 tcaggattcc ctttggactg ggcctggagg tcagaggtca gggaggggcc ccccacctgt
     1441 tcatggacag tggcattgtt tttctcctcg ccatcgtcat agaccgtgta ctcatcctcc
     1501 ggcatagtga acacatcccc gcgagtcacc atatgtatat ctccttctta aagttaaaca
     1561 aaattatttc tagagggaaa ccgttgtggt ctccctatag tgagtcgtat taatttcgcg
     1621 ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     1681 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     1741 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     1801 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
     1861 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc
     1921 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     1981 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     2041 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     2101 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     2161 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     2221 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     2281 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     2341 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc
     2401 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     2461 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     2521 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     2581 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     2641 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     2701 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     2761 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     2821 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     2881 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
     2941 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
     3001 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
     3061 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
     3121 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     3181 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     3241 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     3301 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     3361 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3421 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     3481 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     3541 ctgagagtgc accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3601 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4201 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4381 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4441 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4501 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4561 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4621 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4681 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4741 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4801 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4861 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4921 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4981 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5041 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5101 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5161 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5221 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5281 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5341 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5401 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5461 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5521 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5581 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga a
//
