LOCUS       Exported                9167 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  expresses AmTrac in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    pDR-AmTrac
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9167)
  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. Print 2013.
  PUBMED    23840931
REFERENCE   2  (bases 1 to 9167)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9167
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             772..1023
                     /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     960..981
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1087..1108)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1348..1367)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2279..2303)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2365..2384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2366..2384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2532..2718
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2587..2611)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2730..2747)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2730..2746)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2739..2761)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3182..4528
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4639..5442)
                     /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(5443..5658)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5683..5701)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5769..5873
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5874..6734
                     /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(6092..6111)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6905..7493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7394..7413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7647..7664
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7781..7802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7817..7847
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7855..7871
                     /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     7860..7882
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7879..7895
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7879..7895
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(7925..8380)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8062..8083)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8274..8293
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 actagtgaat tcgcggccat cacaagtttg tacaaaaaag caggctccat gtcaggagca
       61 ataacatgct ctgcggccga tctcgccacc ctacttggcc ccaacgccac ggcggcggcc
      121 gactacattt gcggccaatt aggcaccgtt aacaacaagt tcaccgatgc agccttcgcc
      181 atagacaaca cctacctcct cttctctgcc taccttgtct tcgccatgca gctcggcttc
      241 gctatgcttt gtgctggttc tgttagagcc aagaatacga tgaacatcat gcttaccaat
      301 gtccttgacg ctgcagccgg aggactcttc tactatctct ttggttacgc ctttgccttt
      361 ggaggatcct ccgaagggtt cattggaaga cacaactttg ctcttagaga ctttccgact
      421 cccacagctg attactcttt cttcctctac caatgggcgt tcgcaatcgc ggccgctgga
      481 atcacaagtg gttcgatcgc agagaggact cagttcgtgg cttacttgat atactcttct
      541 ttcttaaccg gatttgttta cccggttgtc tctcactggt tttggtcccc ggatggatgg
      601 gccagtccct ttcgttcagc ggatgatcgt ttgtttagca ccggagccat tgactttgct
      661 ggctccggtg ttgttcacat ggttggtggc atagcaggtt tatggggtgc tcttattgaa
      721 ggtcctcgtc gtggtcggtt cgagaaactc gagaacgtct atatcaaggc cgacaagcag
      781 aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
      841 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      901 aaccactacc tgagcgtcca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
      961 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1021 aagggtggta ccggtggatc tatggtgagc aagggcgagg agctgttcac cggggtggtg
     1081 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1141 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1201 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1261 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1321 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1381 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1441 gacggcaaca tcctggggca caagctggag tacaacttta atggtggtcg cgctattgct
     1501 ctgcgcggcc actctgcctc gctagtagtc ttaggaacct tcctcctatg gtttggatgg
     1561 tatggtttca accccggttc cttcactaag atactcgttc cgtataattc tggttccaac
     1621 tacggccaat ggagcggaat cggccgtaca gcggttaaca ccacactctc aggatgcaca
     1681 gcagctctaa ccacactctt tggtaaacgt ctcctatcag gccactggaa cgtaacggac
     1741 gtttgcaacg ggttactcgg tgggtttgcg gccataaccg caggttgctc cgtcgtagag
     1801 ccatgggcag cgattgtgtg cggcttcatg gcttctgtcg tccttatcgg atgcaacaag
     1861 ctcgcggagc ttgtacaata tgatgatcca ctcgaggcag cccaactaca tggagggtgt
     1921 ggcgcgtggg ggttgatatt cgtaggattg tttgccaaag agaagtatct aaacgaggtt
     1981 tatggcgcca ccccgggaag gccatatgga ctatttatgg gcggaggagg gaagctgttg
     2041 ggagcacaat tggttcaaat acttgtgatt gtaggatggg ttagtgccac aatgggaaca
     2101 ctcttcttca tcctcaaaag gctcaatctg cttaggatct cggagcagca tgaaatgcaa
     2161 gggatggata tgacacgtca cggtggcttt gcttatatct accatgataa tgatgatgag
     2221 tctcatagag tggatcctgg atctcctttc cctcgatcag ctactcctcc tcgcgttgac
     2281 ccagctttct tgtacaaagt ggtgtagtta attcatgatg gccgctgcag gtcgacctcg
     2341 agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg gatccagctt
     2401 tggacttctt cgccagaggt ttggtcaagt ctccaatcaa ggttgtcggc ttgtctacct
     2461 tgccagaaat ttacgaaaag atggaaaagg gtcaaatcgt tggtagatac gttgttgaca
     2521 cttctaaata agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa
     2581 aaataagtgt atacaaattt taaagtgact cttaggtttt aaaacgaaat tcttattctt
     2641 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg tcgctcttat
     2701 tgaccacacc tctaccggca tgccaattca ctggccgtcg ttttacaacg tcgtgactgg
     2761 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2821 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2881 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2941 atcggatcgt acttgttacc catcattgaa ttttgaacat ccgaacctgg gagttttccc
     3001 tgaaacagat agtatatttg aacctgtata ataatatata gtctagcgct ttacggaaga
     3061 caatgtatgt atttcggttc ctggagaaac tattgcatct attgcatagg taatcttgca
     3121 cgtcgcatcc ccggttcatt ttctgcgttt ccatcttgca cttcaatagc atatctttgt
     3181 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
     3241 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
     3301 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
     3361 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
     3421 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
     3481 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
     3541 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
     3601 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
     3661 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
     3721 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
     3781 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
     3841 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
     3901 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
     3961 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
     4021 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
     4081 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
     4141 ctctgaagtt cctatacttt ctagctagag aataggaact tcggaatagg aacttcaaag
     4201 cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg
     4261 ttcacgtcgc acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca
     4321 tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt
     4381 agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag
     4441 cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca
     4501 atgctatcat ttcctttgat attggatcga tccgatgata agctgtcaaa catgagaatt
     4561 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt
     4621 tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc
     4681 tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct
     4741 tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg
     4801 acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta
     4861 accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc
     4921 gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca
     4981 tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg
     5041 cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca
     5101 ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc
     5161 aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt
     5221 aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat
     5281 tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga
     5341 agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact
     5401 aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcct
     5461 gcatgttttt gttctgtgca gttgggttaa gaatactggg caatttcatg tttcttcaac
     5521 actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg
     5581 ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa
     5641 aaaaaatgat gaattgaaaa gctaattctt gaagacgaaa gggcctcgtg atacgcctat
     5701 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5761 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5821 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5881 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5941 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6001 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6061 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6121 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6181 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6241 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6301 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6361 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6421 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6481 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6541 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6601 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6661 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6721 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6781 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6841 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6901 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6961 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7021 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7081 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7141 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7201 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7261 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7321 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7381 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7441 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7501 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7561 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7621 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7681 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7741 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7801 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7861 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cttaccgcat
     7921 caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     7981 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     8041 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     8101 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     8161 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     8221 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     8281 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     8341 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca agcttcctga
     8401 aacggagaaa cataaacagg cattgctggg atcacccata catcactctg ttttgcctga
     8461 ccttttccgg taatttgaaa acaaacccgg tctcgaagcg gagatccggc gataattacc
     8521 gcagaaataa acccatacac gagacgtaga accagccgca catggccgga gaaactcctg
     8581 cgagaatttc gtaaactcgc gcgcattgca tctgtatttc ctaatgcggc acttccaggc
     8641 ctcgatcgag accgtttatc cattgctttt ttgttgtctt tttccctcgt tcacagaaag
     8701 tctgaagaag ctatagtaga actatgagct ttttttgttt ctgttttcct tttttttttt
     8761 tttacctctg tggaaattgt tactctcaca ctctttagtt cgtttgtttg ttttgtttat
     8821 tccaattatg accggtgacg aaacgtggtc gatggtgggt accgcttatg ctcccctcca
     8881 ttagtttcga ttatataaaa aggccaaata ttgtattatt ttcaaatgtc ctatcattat
     8941 cgtctaacat ctaatttctc ttaaattttt tctctttctt tcctataaca ccaatagtga
     9001 aaatcttttt ttcttctata tctacaaaaa cttttttttt ctatcaacct cgttgataaa
     9061 ttttttcttt aacaatcgtt aataattaat taattggaaa ataaccattt tttctctctt
     9121 ttatacacac attcaaaaga aagaaaaaaa atatacccca gcctcga
//
