LOCUS       sequence-7660-ex        5038 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 5038)
  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 5038)
  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: pTK204
FEATURES             Location/Qualifiers
     source          1..5038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(388..406)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      402..449
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      546..1001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(633..652)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     843..864
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1082..1186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1187..1255,1256..2047)
                     /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      2218..2806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2707..2726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2960..2977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3061..3080
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3061..3079
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3109..3131
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3151..3168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3172..3204
                     /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(3208..3216,3217..3231)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(3244..3246,3247..3249,3250..3762)
                     /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    3943..3967
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4292..4316)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(4324..4326,4327..5037)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
ORIGIN
        1 agcttatggc ccaacaatca ccctattcag cagcgatggc agaacagcgt caccaggagt
       61 ggttacgttt tgtcgacctg cttaagaatg cctaccaaaa cgatctccat ttaccgttgt
      121 taaacctgat gctgacgcca gatgagcgcg aagcgttggg gactcgcgtg cgtattgtcg
      181 aagagctgtt gcgcggcgaa atgagccagc gtgagttaaa aaatgaactc ggcgcaggca
      241 tcgcgacgat tacgcgtgga tctaacagcc tgaaagccgc gcccgtcgag ctgcgccagt
      301 ggctggaaga ggtgttgctg aaaagcgatt gaaagcttga tccggctgct aacaaagccc
      361 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
      421 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatctggcg
      481 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
      541 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      721 atttagagct ttacggcacc tcgaccgcaa aaaacttgat ttgggtgatg gttcacgtag
      781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atcgcggtct attcttttga
      901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaat
      961 atttaacgcg aattttaaca aaatattaac gtttacaatt tcgcctgatg cggtattttc
     1021 tccttacgca tctgtgcggt atttcacacc gcatacaggt ggcacttttc ggggaaatgt
     1081 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1141 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1201 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1261 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1321 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1381 aatgatgagc acttttaaag ttctgctatg tgatacacta ttatcccgta ttgacgccgg
     1441 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1501 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1561 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1621 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1681 ggagctgaat gaagccatac caaacgacga gagtgacacc acgatgcctg tagcaatgcc
     1741 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     1801 aatagactga atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     1861 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     1921 agcactgggg ccagatggta agcgctcccg tatcgtagtt atctacacga cggggagtca
     1981 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2041 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2101 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2161 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2221 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2281 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2341 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2401 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2461 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2521 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2581 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2641 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2701 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2761 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2821 ggccttttta cggttcctgg gcttttgctg gccttttgct cacatgttct ttcctgcgtt
     2881 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     2941 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3001 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agatctcgat cccgcgaaat
     3061 taatacgact cactataggg agaccacaac ggtttccctc tagataattt tgtttaactt
     3121 taagaaggag atatacatat gcggggttct catcatcatc atcatcatgg tatggctagc
     3181 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgggc
     3241 cgcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     3301 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     3361 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     3421 accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg
     3481 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     3541 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     3601 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     3661 cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag
     3721 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     3781 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     3841 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     3901 gtcctgctgg agttcgtgac cgccgccggg atcactgata tcacaagttt gtacaaaaaa
     3961 gctgaacgag cccaacaatc accctattca gcagcgatgg cagaacagcg tcaccaggag
     4021 tggttacgtt ttgtcgacct gcttaagaat gcctaccaaa acgatctcca tttaccgttg
     4081 ttaaacctga tgctgacgcc agatgagcgc gaagcgttgg ggactcgcgt gcgtattgtc
     4141 gaagagctgt tgcgcggcga aatgagccag cgtgagttaa aaaatgaact cggcgcaggc
     4201 atcgcgacga ttacgcgtgg atctaacagc ctgaaagccg cgcccgtcga gctgcgccag
     4261 tggctggaag aggtgttgct gaaaagcgat aacccagctt tcttgtacaa agtggtgata
     4321 tcggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4381 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4441 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4501 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
     4561 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4621 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4681 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4741 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     4801 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     4861 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     4921 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4981 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaga
//
