LOCUS       Exported                5479 bp DNA     circular SYN 10-AUG-2021
DEFINITION  Plasmid for chromosomal tagging of Stv1 with iGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5479)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
COMMENT     Linearize with EcoRI for pop-in integration at STV1, then select for
            pop-outs on 5-FOA.
FEATURES             Location/Qualifiers
     source          1..5479
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     CDS             245..694
                     /codon_start=1
                     /label=STV1
                     /translation="GMIISDVASVADSINESVGGGEQGPFNFGDVMIHQVIHTIEFCLN
                     CISHTASYLRLWALSLAHAQLSSVLWDMTISNAFSSKNSGSPLAVMKVVFLFAMWFVLT
                     VCILVFMEGTSAMLHALRLHWVEAMSKFFEGEGYAYEPFSFRAIIE"
     CDS             695..739
                     /codon_start=1
                     /label=Linker
                     /translation="ADPGSSGSSGSSGSS"
     CDS             740..1454
                     /codon_start=1
                     /product="inert GFP variant"
                     /label=iGFP
                     /note="monomeric due to the L221K mutation"
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITHGSSGSSG"
     misc_feature    1466..1965
                     /label=3' STV1
     CDS             complement(2456..3259)
                     /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(3260..3475)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        3573..3677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3678..4538
                     /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      4709..5297
                     /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 atgcgggatg atcatcagtg atgttgccag tgttgcagac agtatcaatg aaagcgttgg
      301 tggaggtgaa caggggccat ttaatttcgg cgatgtcatg attcatcagg taatccatac
      361 tattgaattc tgtttaaatt gtatctctca tacagcatca tatctacgtc tttgggcact
      421 atcgttggcc catgcgcaat tatctagtgt tttatgggat atgacaattt caaacgcttt
      481 cagttctaaa aactcagggt cgcctttggc tgtcatgaag gtggttttct tatttgctat
      541 gtggtttgtt ttaaccgttt gtatcctagt tttcatggaa ggtacttccg caatgttaca
      601 tgcgctacgt ttgcattggg tggaggcaat gtccaaattt tttgaaggtg aaggatatgc
      661 gtacgagcca ttttcatttc gtgcaataat agaagcggat ccaggaagtt cggggtccag
      721 cggttcttca ggcagttcca tggatagcac tgagagcctt ttcactggag ttgtcccaat
      781 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      841 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      901 tgttccttgg ccaacacttg tcactacttt aacttatggt gttcagtgct tttcaagata
      961 cccggatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     1021 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
     1081 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     1141 aaacattctt gggcacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     1201 agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg
     1261 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     1321 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     1381 gagagaccac atggtcctta aagagtttgt aacagctgct gggattacac atggcagctc
     1441 cggctccagc gggtaaagcg gccgctaacc ttaaattact gtaaattttc tcatctaccc
     1501 actgcttttt tcgttttatt tttttttaca catacagaca tataaacgtt gtagaattaa
     1561 acgataaaat ataatatttc ttcacatagt ttacggtata tatatgtctg ggtcggtcaa
     1621 gtggtctacc aataggtcat aaatttgagc agggatctta gcgatgaacc cgacgcccag
     1681 ccgaatgatc tcgtcagcat caaaatccgg aaactgtcgt agtaaattaa caggggaatc
     1741 cgatttgaaa accttggatc tcattttaac taggaatgcc accacaaaga gaatattcat
     1801 atggaatcca tatgcaaaca tgaaatccca cagtttaatg acctgatcaa gcggtttatt
     1861 gcagctggat aacgtgagta tggaaggcat accataaatc tctgctgtaa gtaaattgtc
     1921 cgatagaaat ttactcagct ttggatcaat gattctgaga gaaatcaatt cactggccgt
     1981 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     2041 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2101 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     2161 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     2221 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2281 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2341 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcggg taataactga tataattaaa
     2401 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     2461 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     2521 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     2581 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     2641 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     2701 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     2761 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     2821 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     2881 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     2941 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     3001 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     3061 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     3121 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     3181 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     3241 tatatgtagc tttcgacatg atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt
     3301 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     3361 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     3421 aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa
     3481 aagctggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     3541 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3601 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3661 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3721 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3781 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3841 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3901 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3961 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4021 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4081 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4141 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4201 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4261 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4321 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4381 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4441 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4501 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4561 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4621 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4681 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4741 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4801 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4861 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4921 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4981 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5041 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5101 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5161 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5221 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5281 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5341 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5401 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5461 tgagcgagga agcggaaga
//
