LOCUS       sequence-7661-ex        5059 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 5059)
  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 5059)
  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: pTK234
FEATURES             Location/Qualifiers
     source          1..5059
                     /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(1601..1625)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1633..1635,1636..2349)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2405..2423)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2419..2466
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2563..3018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2650..2669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2860..2881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3099..3203
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3204..3272,3273..4064)
                     /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      4235..4823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4724..4743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4977..4994
                     /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 gtggtgttgc tgaaaagcga attcggcggc
     1261 ggcggcggcg gcggcatggc ccaacaatca ccctattcag cagcgatggc agaacagcgt
     1321 caccaggagt ggttacgttt tgtcgacctg cttaagaatg cctaccaaaa cgatctccat
     1381 ttaccgttgt taaacctgat gctgacgcca gatgagcgcg aagcgttggg gactcgcgtg
     1441 cgtattgtcg aagagctgtt gcgcggcgaa atgagccagc gtgagttaaa aaatgaactc
     1501 ggcgcaggca tcgcgacgat tacgcgtgga tctaacagcc tgaaagccgc gcccgtcgag
     1561 ctgcgccagt ggctggaaga ggtgttgctg aaaagcgaat tcccagcttt cttgtacaaa
     1621 gtggtgatat cggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1681 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1741 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1801 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
     1861 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1921 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1981 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2041 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2101 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2161 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2221 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2281 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2341 tacaagtaaa agcttgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     2401 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     2461 tttttgctga aaggaggaac tatatccgga tctggcgtaa tagcgaagag gcccgcaccg
     2521 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     2581 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2641 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2701 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagagcttta cggcacctcg
     2761 accgcaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2821 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2881 gaacaacact caaccctatc gcggtctatt cttttgattt ataagggatt ttgccgattt
     2941 cggcctattg gttaaaaaat gagctgattt aacaaatatt taacgcgaat tttaacaaaa
     3001 tattaacgtt tacaatttcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3061 tcacaccgca tacaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3121 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3181 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3241 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3301 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3361 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3421 tgctatgtga tacactatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3481 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3541 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3601 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3661 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3721 acgacgagag tgacaccacg atgcctgtag caatgccaac aacgttgcgc aaactattaa
     3781 ctggcgaact acttactcta gcttcccggc aacaattaat agactgaatg gaggcggata
     3841 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3901 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3961 gctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4021 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4081 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4141 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4201 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4261 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4321 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4381 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4441 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4501 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4561 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4621 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4681 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4741 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4801 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctgggct
     4861 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4921 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4981 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5041 ggccgattca ttaatgcag
//
