LOCUS       sequence-359631-        5314 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 5314)
  AUTHORS   Hwang PM, Bunz F, Yu J, Rago C, Chan TA, Murphy MP, Kelso GF, Smith 
            RA, Kinzler KW, Vogelstein B
  TITLE     Ferredoxin reductase affects p53-dependent, 5-fluorouracil-induced 
            apoptosis in colorectal cancer cells.
  JOURNAL   Nat Med. 2001 Oct . 7(10):1111-7.
  PUBMED    11590433
REFERENCE   2  (bases 1 to 5314)
  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 Med. 
            2001 Oct . 7(10):1111-7."
COMMENT     Sequence Label: pTRE-FDXR wt
FEATURES             Location/Qualifiers
     source          1..5314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..30
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    58..76
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    100..118
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    142..160
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    184..202
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    268..286
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        319..357
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     354..378
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      447..456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1941..1950
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1947..1949,1950..1952,1953..2666)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1947..1949,1950..1952,1953..2663)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1992..2013)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2253..2272)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2600..2621
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2680..2814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2704..2723)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2758..2777
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3250..3269)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3363..3380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3534..4122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3614..3633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4293..5084,5085..5153))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4916..4935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5154..5258)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taacctataa aaataggcgt atcacgaggc cctttcgtct tcactcgagt ttaccactcc
       61 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      121 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      181 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      241 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      301 gctcggtacc cgggtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
      361 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      421 caccgggacc gatccagcct ccgcgggcca ccatggcttc gcgctgctgg cgctggtggg
      481 gctggtcggc gtggcctcgg acccggctgc ctcccgccgg gagcaccccg agcttctgcc
      541 accatttctc cacacaggag aagacccccc agatctgtgt ggtgggcagt ggcccagctg
      601 gcttctacac ggcccaacac ctgctaaagc acccccaggc ccacgtggac atctacgaga
      661 aacagcctgt gccctttggc ctggtgcgct ttggtgtggc gcctgatcac cccgaggtga
      721 agaatgtcat caacacattt acccagacgg cccattctgg ccgctgtgcc ttctggggca
      781 acgtggaggt gggcagggac gtgacggtgc cggagctgcg ggaggcctac cacgctgtgg
      841 tgctgagcta cggggcagag gaccatcggg ccctggaaat tcctggtgag gagctgccag
      901 gtgtgtgctc cgcccgggcc ttcgtgggct ggtacaacgg gcttcctgag aaccaggagc
      961 tggagccaga cctgagctgt gacacagccg tgattctggg gcaggggaac gtggctctgg
     1021 acgtggcccg catcctactg accccacctg agcacctgga gagaacggac atcacgaagg
     1081 cagccctggg tgtactgagg cagagtcgag tgaagacagt gtggctagtg ggccggcgtg
     1141 gacccctgca agtggccttc accattaagg agcttcggga gatgattcag ttaccgggag
     1201 cccggcccat tttggatcct gtggatttct tgggtctcca ggacaagatc aaggaggtcc
     1261 cccgcccgag gaagcggctg acggaactgc tgcttcgaac ggccacagag aagccagggc
     1321 cggcggaagc tgcccgccag gcatcggcct cccgtgcctg gggcctccgc tttttccgaa
     1381 gcccccagca ggtgctgccc tcaccagatg ggcggcgggc agcaggtgtc cgcctagcag
     1441 tcactagact ggagggtgtc gatgaggcca cccgtgcagt gcccatggga gacatggaag
     1501 acctcccttg tgggctggtg ctcagcagca ttgggtataa gagccgccct gtcgacccaa
     1561 gcgtgccctt tgactccaag cttggggtca tccccaatgt ggagggccgg gttatggatg
     1621 tgccaggcct ctactgcagc ggctgggtga agagaggacc tacaggtgtc atagccacaa
     1681 ccatgactga cagcttcctc accggccaga tgctgctgca ggacctgaag gctgggttgc
     1741 tcccctctgg ccccaggcct ggctacgcag ccatccaggc cctgctcagc agccgagggg
     1801 tccggccagt ctctttctca gactgggaga agctggatgc cgaggaggtg gcccggggcc
     1861 agggcacggg gaagcccagg gagaagctgg tggatcctca ggagatgctg cgcctcctgg
     1921 gccacggagt cggaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1981 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2041 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2101 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2161 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2221 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2281 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2341 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2401 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2461 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2521 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2581 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2641 tcggcatgga cgagctgtac aagtaaagcg gccctagagg atccagacat gataagatac
     2701 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2761 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2821 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2881 aagtaaaacc tctacaaatg tggtatggct gattatgatc ctgcaagcct cgtcgtcctg
     2941 gccggaccac gctatctgtg caaggtcccc ggccccggac gcgcgctcca tgagcagagc
     3001 gcccgccgcc gaggcgaaga ctcgggcggc gccctgcccg tcccaccagg tcaacaggcg
     3061 gtaaccggcc tcttcatcgg gaatgcgcgc gaccttcagc atcgccggca tgtccccctg
     3121 gcggacggga agtatccagc tcgaccaagc ttggcgagat tttcaggagc taaggaagct
     3181 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     3241 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     3301 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3361 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3421 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3481 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3541 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3601 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3661 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3721 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3781 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3841 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3901 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3961 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4021 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4081 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4141 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4201 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4261 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4321 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4381 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4441 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4501 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4561 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4621 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4681 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4741 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4801 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4861 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     4921 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4981 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5041 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5101 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5161 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5221 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5281 cctgacgtct aagaaaccat tattatcatg acat
//
