LOCUS       sequence-68721-e        9174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses MepTrac-H194E in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9174)
  AUTHORS   De Michele R, Ast C, Loque D, Ho CH, Andrade SL, Lanquar V, 
            Grossmann G, Gehne S, Kumke MU, Frommer WB
  TITLE     Fluorescent sensors reporting the activity of ammonium transceptors 
            in live cells.
  JOURNAL   eLife. 2013 Jul 2;2:e00800.
  PUBMED    23840931
REFERENCE   2  (bases 1 to 9174)
  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: "eLife."; 
            date: "2013-07-2"; volume: "2"; pages: "e00800"
COMMENT     Sequence Label: pDR-meptrac-H194E
FEATURES             Location/Qualifiers
     source          1..9174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..46
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             724..975
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNH
                     YLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     912..933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1039..1060)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1300..1319)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2286..2310)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2372..2391)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2373..2391)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2539..2725
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2594..2618)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2737..2754)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2737..2753)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2746..2768)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3189..4535
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4646..5449)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5450..5665)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5690..5708)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5776..5880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5881..5949,5950..6741)
                     /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(6099..6118)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6912..7500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7401..7420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7654..7671
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7788..7809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7824..7829,7830..7847,7848..7854)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7862..7878
                     /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)."
     primer_bind     7867..7889
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7886..7902
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7886..7902
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7932..8387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8069..8090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8281..8300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 actagtgaat tcgcggccat cacaagtttg tacaaaaaag caggcttaat gtcttacaat
       61 tttacaggta cgcctacagg cgaaggaacg ggtggtaact cgttgacaac agatttgaat
      121 acacaatttg acttggccaa catgggatgg atcggtgtgg cttcagcagg tgtgtggatt
      181 atggtcccag gtatcggttt attatattct ggtttatcca ggaaaaagca tgctttatcc
      241 ttgctttggg cctcgatgat ggcttccgcc gtgtgtattt tccaatggtt tttctgggga
      301 tactcattag ctttctcaca caacactaga ggtaacggtt ttattggtac cttggaattc
      361 tttgggtttc gtaacgtttt aggagcccca tctagtgtca gttctcttcc cgatatactg
      421 tttgccgttt accaaggtat gtttgccgca gtcaccggtg ccctaatgct aggtggtgcc
      481 tgcgagaggg caaggttgtt tcctatgatg gtgttcttgt ttttatggat gactattgtt
      541 tattgtccta ttgcatgctg ggtctggaat gccgagggtt ggttggtcaa attgggtagc
      601 ttggactatg caggtggttt atgtgtcgag ttaacatctg gacatggtgg tctagtttac
      661 gctttgatac tgggtaagcg taatgaccct gttacacgtt tgtctaacgt ctatatcaag
      721 gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac
      781 ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga cggccccgtg
      841 ctgctgcccg acaaccacta cctgagcgtc cagtccaagc tgagcaaaga ccccaacgag
      901 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      961 gacgagctgt acaagggtgg taccggtgga tctatggtga gcaagggcga ggagctgttc
     1021 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1081 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1141 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1201 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1261 cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1321 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1381 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt taataaaggg
     1441 atgcccaagt acaaaccaca ttccgtcacc tcggtggttt taggcacagt gttcttatgg
     1501 tttggttgga tgttctttaa cggaggctct gcaggtaatg caactatacg agcatggtac
     1561 tctattatgt ccacaaactt agctgctgct tgcggtggct tgacatggat ggttatcgat
     1621 tatttcagat gcggaagaaa gtggactaca gttggtttgt gttcaggtat catcgctggc
     1681 ctagtgggta tcaccccagc cgccgggttc gtgccaatct ggtcagccgt tgtcattggt
     1741 gtggttactg gtgcaggatg taaccttgct gttgacttaa agagtctatt gcgcatcgat
     1801 gatggtctag attgttactc tatccatggt gtgggtggtt gtattggttc tgtattaact
     1861 ggtatctttg ctgcagacta tgtaaatgcc actgcaggct cttacattag tccaattgat
     1921 ggtggctgga tcaatcatca ctataaacaa gttggttatc aattagcagg tatatgcgct
     1981 gcactagcct ggactgttac tgtcacatct atcttgcttc taactatgaa tgccattcca
     2041 tttttaaaac taagattaag tgctgatgag gaagaattag gtaccgacgc tgctcaaatt
     2101 ggtgaattta catacgagga atccactgct tacatcccag aaccaatcag atctaaaaca
     2161 tcggcacaaa tgccacctcc tcatgaaaac attgatgata agattgtggg taacacagac
     2221 gcagaaaaga attctacgcc ttccgacgct tcttctacta agaacactga ccatatagta
     2281 taaagaccca gctttcttgt acaaagtggt gtagttaatt catgatggcc gctgcaggtc
     2341 gacctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt acgcgcggat
     2401 ccagctttgg acttcttcgc cagaggtttg gtcaagtctc caatcaaggt tgtcggcttg
     2461 tctaccttgc cagaaattta cgaaaagatg gaaaagggtc aaatcgttgg tagatacgtt
     2521 gttgacactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
     2581 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaattct
     2641 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg
     2701 ctcttattga ccacacctct accggcatgc caattcactg gccgtcgttt tacaacgtcg
     2761 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     2821 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2881 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2941 ccgcataatc ggatcgtact tgttacccat cattgaattt tgaacatccg aacctgggag
     3001 ttttccctga aacagatagt atatttgaac ctgtataata atatatagtc tagcgcttta
     3061 cggaagacaa tgtatgtatt tcggttcctg gagaaactat tgcatctatt gcataggtaa
     3121 tcttgcacgt cgcatccccg gttcattttc tgcgtttcca tcttgcactt caatagcata
     3181 tctttgttaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc
     3241 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag
     3301 cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag
     3361 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     3421 gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc
     3481 ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct
     3541 ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc
     3601 tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact
     3661 gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct
     3721 ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca
     3781 ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa
     3841 tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt
     3901 tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa
     3961 gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag
     4021 caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg
     4081 ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct tttggttttc
     4141 aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcg gaataggaac
     4201 ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa cgcgagctgc gcacatacag
     4261 ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt atatatatat acatgagaag
     4321 aacggcatag tgcgtgttta tgcttaaatg cgtacttata tgcgtctatt tatgtaggat
     4381 gaaaggtagt ctagtacctc ctgtgatatt atcccattcc atgcggggta tcgtatgctt
     4441 ccttcagcac taccctttag ctgttctata tgctgccact cctcaattgg attagtctca
     4501 tccttcaatg ctatcatttc ctttgatatt ggatcgatcc gatgataagc tgtcaaacat
     4561 gagaattggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     4621 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     4681 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     4741 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     4801 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     4861 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     4921 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     4981 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     5041 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     5101 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     5161 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     5221 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     5281 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     5341 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     5401 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     5461 gtttcctgca tgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     5521 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     5581 ccttctgttc ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag
     5641 aataaaaaaa aaatgatgaa ttgaaaagct aattcttgaa gacgaaaggg cctcgtgata
     5701 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5761 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5821 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5881 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5941 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6001 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6061 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6121 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6181 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6241 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6301 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6361 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6421 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6481 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6541 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6601 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6661 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6721 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6781 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6841 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6901 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6961 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7021 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7081 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7141 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7201 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7261 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7321 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7381 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7441 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7501 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7561 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7621 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7681 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     7741 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7801 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7861 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
     7921 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     7981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     8041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     8101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     8161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     8221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     8281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     8341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccaagc
     8401 ttcctgaaac ggagaaacat aaacaggcat tgctgggatc acccatacat cactctgttt
     8461 tgcctgacct tttccggtaa tttgaaaaca aacccggtct cgaagcggag atccggcgat
     8521 aattaccgca gaaataaacc catacacgag acgtagaacc agccgcacat ggccggagaa
     8581 actcctgcga gaatttcgta aactcgcgcg cattgcatct gtatttccta atgcggcact
     8641 tccaggcctc gatcgagacc gtttatccat tgcttttttg ttgtcttttt ccctcgttca
     8701 cagaaagtct gaagaagcta tagtagaact atgagctttt tttgtttctg ttttcctttt
     8761 tttttttttt acctctgtgg aaattgttac tctcacactc tttagttcgt ttgtttgttt
     8821 tgtttattcc aattatgacc ggtgacgaaa cgtggtcgat ggtgggtacc gcttatgctc
     8881 ccctccatta gtttcgatta tataaaaagg ccaaatattg tattattttc aaatgtccta
     8941 tcattatcgt ctaacatcta atttctctta aattttttct ctttctttcc tataacacca
     9001 atagtgaaaa tctttttttc ttctatatct acaaaaactt tttttttcta tcaacctcgt
     9061 tgataaattt tttctttaac aatcgttaat aattaattaa ttggaaaata accatttttt
     9121 ctctctttta tacacacatt caaaagaaag aaaaaaaata taccccagcc tcga
//
