LOCUS       sequence-7658-ex        7585 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 7585)
  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 7585)
  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: pTK222
FEATURES             Location/Qualifiers
     source          1..7585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(388..406)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     primer_bind     complement(443..460)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    449..673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      719..1147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(806..825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1016..1037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1156..1176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1161..1490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1341..1476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1403..1422
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1557..2351
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1611..1630)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2221..2240
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2525..2646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2562..2581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2616..2635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2695..2711)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2695..2711)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2708..2730)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2719..2735
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2743..2749,2750..2767,2768..2773))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2788..2809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2926..2943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3097..3682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3177..3196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3853..4644,4645..4713))
                     /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     4476..4495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4714..4818)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4893..4912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5084..5463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5464..5667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5618..5638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5712..5731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5712..5730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5791..5793,5794..5796,5797..6309)
                     /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    6490..6514
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6839..6863)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(6871..6873,6874..7584)
                     /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 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
      481 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
      541 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
      601 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
      661 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
      721 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
      781 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
      841 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
      901 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
      961 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1021 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1081 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1141 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1321 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1381 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     1441 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     1501 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     1561 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     1621 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     1681 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     1741 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     1801 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     1861 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     1921 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     1981 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2041 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     2101 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2161 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2221 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2281 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2341 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     2401 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     2461 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     2521 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2581 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2641 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     2701 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2761 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2821 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2881 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2941 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3001 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3061 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3121 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3181 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3241 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3301 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3361 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3421 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3481 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3541 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3601 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     3661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4141 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4441 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     4861 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4921 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     4981 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     5041 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     5101 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5161 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5221 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5281 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5341 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5401 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5461 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5521 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5581 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5641 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
     5701 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta
     5761 agcttggtac cgagctcgga tccgggccgc atggtgagca agggcgagga gctgttcacc
     5821 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     5881 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     5941 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag
     6001 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     6061 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     6121 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     6181 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacat cagccacaac
     6241 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     6301 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     6361 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     6421 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     6481 actgatatca caagtttgta caaaaaagct gaacgagccc aacaatcacc ctattcagca
     6541 gcgatggcag aacagcgtca ccaggagtgg ttacgttttg tcgacctgct taagaatgcc
     6601 taccaaaacg atctccattt accgttgtta aacctgatgc tgacgccaga tgagcgcgaa
     6661 gcgttgggga ctcgcgtgcg tattgtcgaa gagctgttgc gcggcgaaat gagccagcgt
     6721 gagttaaaaa atgaactcgg cgcaggcatc gcgacgatta cgcgtggatc taacagcctg
     6781 aaagccgcgc ccgtcgagct gcgccagtgg ctggaagagg tgttgctgaa aagcgataac
     6841 ccagctttct tgtacaaagt ggtgatatcg gtgagcaagg gcgaggagct gttcaccggg
     6901 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     6961 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     7021 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc
     7081 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     7141 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     7201 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     7261 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     7321 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     7381 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     7441 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     7501 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     7561 ctcggcatgg acgagctgta caaga
//
