LOCUS       sequence-7659-ex        4708 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 4708)
  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 4708)
  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: pTK164
FEATURES             Location/Qualifiers
     source          1..4708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             67..89
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             109..126
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..162
                     /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(166..174,175..189)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(202..204,205..207,208..720)
                     /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    901..925
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1250..1274)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1282..1284,1285..1998)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2054..2072)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2068..2115
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2212..2667
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2299..2318)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2509..2530
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2748..2852
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2853..2921,2922..3713)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3884..4472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4373..4392
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4626..4643
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gataattttg tttaacttta agaaggagat atacatatgc ggggttctca tcatcatcat
      121 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
      181 gacgataagg atccgggccg catggtgagc aagggcgagg agctgttcac cggggtggtg
      241 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      301 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      361 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
      421 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      481 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      541 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      601 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
      661 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      721 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      781 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      841 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactgatatc
      901 acaagtttgt acaaaaaagc tgaacgagcc caacaatcac cctattcagc agcgatggca
      961 gaacagcgtc accaggagtg gttacgtttt gtcgacctgc ttaagaatgc ctaccaaaac
     1021 gatctccatt taccgttgtt aaacctgatg ctgacgccag atgagcgcga agcgttgggg
     1081 actcgcgtgc gtattgtcga agagctgttg cgcggcgaaa tgagccagcg tgagttaaaa
     1141 aatgaactcg gcgcaggcat cgcgacgatt acgcgtggat ctaacagcct gaaagccgcg
     1201 cccgtcgagc tgcgccagtg gctggaagag gtgttgctga aaagcgataa cccagctttc
     1261 ttgtacaaag tggtgatatc ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1321 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1381 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1441 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     1501 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1561 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1621 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1681 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1741 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1801 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1861 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     1921 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1981 gacgagctgt acaagtaaaa gcttgatccg gctgctaaca aagcccgaaa ggaagctgag
     2041 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2101 ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat agcgaagagg
     2161 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2221 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2281 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2341 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agagctttac
     2401 ggcacctcga ccgcaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     2461 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2521 tccaaactgg aacaacactc aaccctatcg cggtctattc ttttgattta taagggattt
     2581 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaatattt aacgcgaatt
     2641 ttaacaaaat attaacgttt acaatttcgc ctgatgcggt attttctcct tacgcatctg
     2701 tgcggtattt cacaccgcat acaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2761 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2821 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2881 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2941 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3001 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3061 ttaaagttct gctatgtgat acactattat cccgtattga cgccgggcaa gagcaactcg
     3121 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3181 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3241 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3301 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3361 ccataccaaa cgacgagagt gacaccacga tgcctgtagc aatgccaaca acgttgcgca
     3421 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactgaatgg
     3481 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3541 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3601 atggtaagcg ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3661 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3721 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3781 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3841 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3901 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3961 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4021 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4081 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4141 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4201 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4261 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4321 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4381 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4441 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4501 tcctgggctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4561 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4621 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4681 ccgcgcgttg gccgattcat taatgcag
//
