LOCUS       sequence-13879-e        5987 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 5987)
  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 5987)
  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_FL
FEATURES             Location/Qualifiers
     source          1..5987
                     /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             1906..1928
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1959..1978)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1960..1978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2062..2081)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             3539..3730
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3724..3746)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3832..3974
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3887..3906
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3989..4006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4160..4748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4240..4259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4919..5710,5711..5779))
                     /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     5542..5561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5780..5884)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5952..5970
                     /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 gatccctaca gatggccagg agctgggcag
      541 cccagccagt actgagcgat ggagcgtggg tagggagggt ccacagtgtc cactcgccgt
      601 gtgcgaagat tgactcggta gtacttgtct ccagagaaga agaagacact ctggatgggt
      661 tcacaggtgg caggcacaag ccagtccatc ctgtagtcat catagttgtt ggctcccaag
      721 ttgctctcct cactggagaa caaggacagc cacatggcgc gggatggccg gcgggagttc
      781 tggttgcggc cacggctgtg gcctcgttgt gaacggtagc ctttgcggtt gcgatgccta
      841 aacctttgtt tcttggccaa ggaggggcgg ggtgccatgc ctgagatgta gatgcggcca
      901 gccatggctg cgtccacttg ccctggcaca ccgtgccagt cccggctaat gaactggggc
      961 tgtctggtac cagcagaggt tctgccccag aagagaagct cgaagatgtc ctcccagctg
     1021 tcccgctgca tcatggcaaa gtgttcaaac acagccgaca gggagctgcc ttcacactcc
     1081 tcctgactgg gctggtgctg gaactggtac tcccagtact gtttcccctt gaagaagtag
     1141 acccgctccc ggccactgta gctatgggca gggagggcca aggctgcatc cacgttgtcc
     1201 gggatgccat cgaagccgtc agagatattt cgggggtaat cagggtccag gacaccatcc
     1261 tcaaagcgcc agtactgact acccttgaag aggtaggtct tcccctgaca gttgatgcgg
     1321 gtgaaggcgg catcgatggg gccctcgatg ccccagacat ctcggatgag cttggggtac
     1381 ccaggcctca ctgccttttc gtccagttca tagcagtact gccctcggaa ggcaaagagg
     1441 gaaccgttct tgaggtcggt gaaggcgtcg aagggcttcc cactgcacag ctcctcctct
     1501 gctgggggct gaggtctccc tggatgaagg gtctcaggcc ttgagtctat cccctcaggc
     1561 ttagaggcgc ccacctcagg cgcaggggcc tcttcctcag gtttcagaac aggtgtctgc
     1621 tcaggattcc ctttggactg ggcctggagg tcagaggtca gggaggggcc ccccacctgt
     1681 tcatggacag tggcattgtt tttctcctcg ccatcgtcat agaccgtgta ctcatcctcc
     1741 ggcatagtga acacatcccc gcgagtcact tggggcttgc actcagccgt atagtctgtg
     1801 cagcagctct ggtagtaaga gcagagctcg tcacactggc acttcttgtc cacgttgaag
     1861 ccctcagtgc agcggccctt gcatgactct tggtccatat gtatatctcc ttcttaaagt
     1921 taaacaaaat tatttctaga gggaaaccgt tgtggtctcc ctatagtgag tcgtattaat
     1981 ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg
     2041 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     2101 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     2161 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
     2221 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgac
     2281 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     2341 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     2401 cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt
     2461 cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca
     2521 ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct
     2581 acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg
     2641 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg
     2701 accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg
     2761 gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat
     2821 ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga
     2881 gccgggccac ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc
     2941 aagaattgga gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca
     3001 gaacatatcc atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggcagcgt
     3061 tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg
     3121 gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga
     3181 ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt
     3241 ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat
     3301 cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca
     3361 ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc
     3421 acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc
     3481 tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt
     3541 gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac
     3601 gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct
     3661 tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga
     3721 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     3781 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     3841 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     3901 attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     3961 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4021 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4081 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4141 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4561 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4741 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4861 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4921 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4981 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5041 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5101 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5161 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5221 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5281 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5341 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5401 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5461 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5521 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5581 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5641 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5701 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5761 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5821 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5881 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     5941 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaa
//
