LOCUS       sequence-229454-        4983 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 4983)
  AUTHORS   Kaper T, Looger LL, Takanaga H, Platten M, Steinman L, Frommer WB
  TITLE     Nanosensor detection of an immunoregulatory tryptophan 
            influx/kynurenine efflux cycle.
  JOURNAL   PLoS Biol. 2007 Oct;5(10):e257.
  PUBMED    17896864
REFERENCE   2  (bases 1 to 4983)
  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: "PLoS 
            Biol."; date: "2007-10"; volume: "5(10)"; pages: "e257"
COMMENT     Sequence Label: pTK203
FEATURES             Location/Qualifiers
     source          1..4983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     200..219
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        200..218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             250..272
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             292..309
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             313..345
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(349..357,358..372)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(712..714,715..717,718..1230)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     protein_bind    1411..1435
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1760..1784)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1792..1794,1795..2508)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2564..2582)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2578..2625
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2722..3177
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2809..2828)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3019..3040
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3203..3307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3308..3376,3377..4168)
                     /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     complement(3526..3545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4339..4927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4828..4847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      241 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      301 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      361 gatgacgata aggatccggc ccaacaatca ccctattcag cagcgatggc agaacagcgt
      421 caccaggagt ggttacgttt tgtcgacctg cttaagaatg cctaccaaaa cgatctccat
      481 ttaccgttgt taaacctgat gctgacgcca gatgagcgcg aagcgttggg gactcgcgtg
      541 cgtattgtcg aagagctgtt gcgcggcgaa atgagccagc gtgagttaaa aaatgaactc
      601 ggcgcgggca tcgcgacgat tacgcgtgga tctaacagcc tgaaagccgc gcccgttgag
      661 ctgcgccagt ggctggaaga ggtgttgctg aaaagcgagg atccgggccg catggtgagc
      721 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      781 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      841 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      901 accctgacct ggggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
      961 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1021 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1081 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1141 tacaactaca tcagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     1201 gccaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1261 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1321 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1381 ttcgtgaccg ccgccgggat cactgatatc acaagtttgt acaaaaaagc aggctcggcc
     1441 caacaatcac cctattcagc agcgatggca gaacagcgtc accaggagtg gttacgtttt
     1501 gtcgacctgc ttaagaatgc ctaccaaaac gatctccatt taccgttgtt aaacctgatg
     1561 ctgacgccag atgagcgcga agcgttgggg actcgcgtgc gtattgtcga agagctgttg
     1621 cgcggcgaaa tgagccagcg tgagttaaaa aatgaactcg gcgcaggcat cgcgacgatt
     1681 acgcgtggat ctaacagcct gaaagccgcg cccgtcgagc tgcgccagtg gctggaagag
     1741 gtgttgctga aaagcgataa cccagctttc ttgtacaaag tggtgatatc ggtgagcaag
     1801 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1861 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1921 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1981 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2041 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2101 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2161 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2221 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     2281 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
     2341 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2401 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2461 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaaa gcttgatccg
     2521 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     2581 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     2641 atatccggat ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2701 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2761 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2821 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2881 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2941 gtgttggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3001 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3061 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3121 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     3181 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3241 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3301 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3361 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3421 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3481 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3541 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3601 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3661 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3721 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3781 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3841 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3901 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3961 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4021 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4081 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4141 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4201 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4261 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4321 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4381 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4441 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4501 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4561 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4621 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4681 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4741 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4801 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4861 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4921 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4981 tca
//
