LOCUS       Exported                6013 bp DNA     circular SYN 09-AUG-2021
DEFINITION  Plasmid for chromosomal tagging of Nhx1 with iGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
COMMENT     Linearize with BglII for pop-in integration at NHX1, then select for
            pop-outs on 5-FOA.
FEATURES             Location/Qualifiers
     source          1..6013
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     CDS             245..1000
                     /codon_start=1
                     /label=NHX1
                     /translation="AAISICVARWCAVFPLSQFVNWIYRVKTIRSMSGITGENISVPDE
                     IPYNYQMMTFWAGLRGAVGVALALGIQGEYKFTLLATVLVVVVLTVIIFGGTTAGMLEV
                     LNIKTGCISEEDTSDDEFDIEAPRAINLLNGSSIQTDLGPYSDNNSPDISIDQFAVSSN
                     KNLPNNISTTGGNTFGGLNETENTSPNPARSSMDKRNLRDKLGTIFNSDSQWFQNFDEQ
                     VLKPVFLDNVSPSLQDSATQSPADFSSQNH"
     CDS             1010..1045
                     /codon_start=1
                     /label=linker
                     /translation="GSSGSSGSSGSS"
     CDS             1046..1762
                     /codon_start=1
                     /label=iGFP
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITHGSSGSSG"
     misc_feature    1772..2487
                     /label=3' NHX1
     CDS             complement(2990..3793)
                     /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(3794..4014)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        4107..4211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4212..5072
                     /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"
     rep_origin      5243..5831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcgcagct atttctatat gtgttgctcg ttggtgtgct gtgttcccat tgtcgcaatt
      301 tgttaactgg atatatagag taaagacaat cagatctatg agcggcataa ccggagaaaa
      361 tatttctgtt cccgatgaaa taccctacaa ttaccaaatg atgacatttt gggcaggttt
      421 acgtggtgct gttggtgtcg ccttggcgtt gggaattcaa ggtgagtata agttcacttt
      481 attggcaacg gtccttgttg ttgttgtttt aacagttatc atttttgggg gcactactgc
      541 aggaatgtta gaagttttaa atattaagac tggttgcata agtgaagaag atacatctga
      601 tgacgagttt gatatagagg ctccaagggc gataaattta ttgaacggta gttctattca
      661 gacagatttg ggcccatatt ctgacaacaa ttctccagat atttcaattg accaattcgc
      721 ggtcagcagt aacaagaatc tccccaataa catatccaca actggtggta atacttttgg
      781 aggccttaat gaaactgaga atactagccc taacccggca aggtcttcta tggataagcg
      841 taatttgaga gataaactgg gaacaatctt taattccgac tcacaatggt ttcaaaattt
      901 tgatgaacag gtattgaagc cagtattctt ggacaacgtt tctccatcct tacaagattc
      961 ggctacgcaa tcacctgcag atttctcttc ccaaaaccac acggatccag gaagttcggg
     1021 gtccagcggt tcttcaggca gttccatgga tagcactgag agccttttca ctggagttgt
     1081 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     1141 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
     1201 actacctgtt ccttggccaa cacttgtcac tactttaact tatggtgttc agtgcttttc
     1261 aagatacccg gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta
     1321 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
     1381 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
     1441 agatggaaac attcttgggc acaaattgga atacaactat aactcacaca atgtatacat
     1501 catggcagac aaacaaaaga atggaatcaa agctaacttc aaaattagac acaacattga
     1561 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     1621 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
     1681 cgaaaagaga gaccacatgg tccttaaaga gtttgtaaca gctgctggga ttacacatgg
     1741 cagctccggc tccagcgggt aaagcggccg cactttaaag tgtatggttt ccttgtttct
     1801 aatataaata tattaattaa gtctttatac attctctctt actcaacgcc gaaggaatac
     1861 ccttctacac tttattcatt ttaaatcatg tactgttata tacacgctag taggattgat
     1921 cttatttaat catatttcgc acgttacccg actgctttca gttttgccgc tttgtgataa
     1981 tgaggaattt cagctagctt ttaatgagcc gatactaaat caccatgatt ttgggagagg
     2041 aaaagaagaa aaggtaaaac aacgaatatt tttcaaatac cttagttact gaatatccat
     2101 aactactcgt ataattggtg agaaaatagt gcatatttag tttacttttt gcctttgatt
     2161 gaaaatatat attcatggtg ctttttactc ggtgtgaaaa ggcaagaaag gagaaactcg
     2221 ccgctggtta taagccctta gttgattatt tgattgattg tgatactccc acatttttag
     2281 aaagaattga ggcaatccaa gagtgggacc gatccagaga tgatctttat gtttggattc
     2341 ccattctaga cagaatggac gggctgttgt taaaagtggc agagaaatat aaatacaagc
     2401 aagaccccaa gaaggagtgt gaagtcaaat tggtggaaat ggaagcgcat gatgttgatt
     2461 attgtttgaa aatgctaaag tttactcggt accgagctcg aattcactgg ccgtcgtttt
     2521 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2581 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2641 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2701 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2761 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2821 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2881 gtcatcaccg aaacgcgcga gacgaaaggg cgggtaataa ctgatataat taaattgaag
     2941 ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct
     3001 ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct
     3061 tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag
     3121 agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac
     3181 ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt
     3241 gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag
     3301 tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc
     3361 taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca
     3421 ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt
     3481 actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat
     3541 tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca
     3601 agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca
     3661 gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca
     3721 tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg
     3781 tagctttcga catgatttat cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta
     3841 agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa
     3901 gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc
     3961 aaagaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg
     4021 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4081 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4141 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4201 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4261 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4321 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4381 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4441 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4501 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4561 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4621 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4681 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4741 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4801 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4861 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4921 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4981 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5041 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5101 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5161 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5221 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5281 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5341 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5401 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5461 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5521 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5581 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5641 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5701 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5761 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5821 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5881 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5941 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6001 aggaagcgga aga
//
