LOCUS       Exported                5759 bp DNA     circular SYN 10-AUG-2021
DEFINITION  Plasmid for chromosomal tagging of Mnn9 with yeast codon-optimized 
            msGFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5759)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
COMMENT     Linearize with AgeI for pop-in integration at MNN9, then select for 
            pop-outs on 5-FOA.
FEATURES             Location/Qualifiers
     source          1..5759
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     CDS             247..810
                     /codon_start=1
                     /label=MNN9
                     /translation="ERRAAMALARNELLFSTIGPHTSWVLWLDADIIETPPSLIQDMTK
                     HNKAILAANIYQRFYDEEKKQPSIRPYDFNNWQESDTGLEIASQMGDDEIIVEGYAEIA
                     TYRPLMAHFYDANGVPGEEMALDGVGGGCTLVKAEVHRDGAMFPNFPFYHLIETEGFAK
                     MAKRLNYDVFGLPNYLVYHIEEENH"
     CDS             811..855
                     /codon_start=1
                     /label=linker
                     /translation="SDPGSSGSSGSSGAS"
     CDS             856..1563
                     /codon_start=1
                     /label=yo-msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     misc_feature    1578..2233
                     /label=3' MNN9
     CDS             complement(2736..3539)
                     /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(3540..3755)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        3853..3957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3958..4818
                     /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      4989..5577
                     /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 atgcaggaaa gacgtgcagc aatggctttg gcgcgcaatg aattactatt ctccaccata
      301 ggacctcaca cttcttgggt gctgtggcta gatgccgata ttatagagac accaccatct
      361 ttaattcaag acatgaccaa acacaacaaa gctatcttag ctgcaaacat ttatcaaaga
      421 ttttacgatg aagagaagaa gcaaccatca atcagaccat acgatttcaa caactggcaa
      481 gaaagtgaca ccggtttaga aatagcctct cagatgggtg atgacgagat tattgtcgag
      541 ggttatgcag aaattgccac ttataggcca ttaatggctc atttctacga tgctaatggc
      601 gtaccaggtg aagagatggc gctggatggt gttggtggag gctgtacttt ggtcaaagca
      661 gaagttcaca gagacggtgc catgttccct aatttcccat tttatcactt gattgaaaca
      721 gaaggttttg ctaagatggc gaagagatta aactacgatg tatttggctt accaaactat
      781 ttggtttatc acatagagga agagaaccat tcggatccgg gaagttcggg gtccagcggt
      841 tcttcaggcg ctagtatgga tagcactgag agcttgttca caggtgttgt tcctatcttg
      901 gtcgaattag acggtgacgt aaacggtcat aagttcagtg ttagaggtga aggtgaaggt
      961 gacgctacta acggtaaatt gacattgaag tttatttgta ctacaggtaa attgccagtt
     1021 ccttggccaa ctttggttac tacattgaca tacggtgttc aatgtttttc aagataccca
     1081 gatcatatga agcaacatga tttctttaag tctgctatgc cagaaggtta cgttcaagaa
     1141 agaactatca cttttaaaga tgatggtact tacaagacaa gagcagaggt taagttcgaa
     1201 ggtgacactt tggttaacag aatcgaattg aagggtatcg attttaaaga agatggtaat
     1261 attttgggtc ataaattgga atacaactac aactcacata acgtttacat tacagctgat
     1321 aaacaaaaga atggtattaa agcaaacttc aagatcagac ataatgttga agatggttct
     1381 gttcaattag cagatcatta ccaacaaaac actccaattg gtgacggtcc agttttgtta
     1441 ccagataacc attacttgtc tacacaatca aaattgtcta aagatccaaa cgaaaagaga
     1501 gatcatatgg ttttgttaga atttgttact gctgctggta tcacaggtgg cagtggcggt
     1561 tcttaataag cggccgcgca actgagcaaa aagcatacag aagctatagc gttattgaaa
     1621 gataatattg ttgcgattat tagttgaaaa taaacaaaac aagatatata ttcatatata
     1681 tatatatata atttacacat gtatattaaa aaatgccgaa taaaaagtgc tacaacaact
     1741 aatttctgag cttgagaata aaccattcac attttatttc ttttggtgct tctttatcat
     1801 ttataaatcg gtggtcttga ctttttattc ctttctagtc taacaatcat ccattctata
     1861 gcgtctttaa caccttctcc agtcaatgcg cttattggta aaaccctact atctctagcg
     1921 cttatatgtt ccgcaatctt attaaatact tcttttatat cttgtacttc cattctatct
     1981 tgtctatctt gtttattggc cagcatcaag atgggtacac cttcaatttc ttcatccatt
     2041 acaactgact gtagggtcgt ggaacattcg tctaatcgtt ctctatctga actatccaca
     2101 atgaaaatta taccatgaca tagggaatag tattcggacc acattgatct cagtgattct
     2161 tgaccaccta catcccaaaa cttcaagatc tgtttactgt caacgggtat tgtcgccaca
     2221 ttttgtccta ccgggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact
     2281 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2341 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2401 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2461 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2521 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2581 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2641 gcgcgagacg aaagggcggg taataactga tataattaaa ttgaagctct aatttgtgag
     2701 tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc
     2761 aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc
     2821 tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc
     2881 acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc
     2941 ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg
     3001 ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta
     3061 gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt
     3121 acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca
     3181 ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact
     3241 gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat
     3301 aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt
     3361 gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg
     3421 gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc
     3481 ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg
     3541 atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt gggttaagaa tactgggcaa
     3601 tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc
     3661 cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat
     3721 caaaaaaaag aataaaaaaa aaatgatgaa ttgaattgaa aagctggcct cgtgatacgc
     3781 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3841 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3901 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3961 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4021 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4081 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4141 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4201 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4261 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4321 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4381 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4441 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4501 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4561 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4621 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4681 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4741 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4801 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4861 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4921 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4981 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5041 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5101 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5161 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5221 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5281 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5341 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5401 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5461 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5521 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5581 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5641 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5701 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaga
//
