LOCUS       Exported                5528 bp DNA     circular SYN 10-AUG-2021
DEFINITION  Plasmid for chromosomal tagging of Aur1 with yeast codon-optimized 
            msGFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5528)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
COMMENT     Linearize with PstI for pop-in integration at AUR1, then select for 
            pop-outs on 5-FOA.
FEATURES             Location/Qualifiers
     source          1..5528
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     CDS             239..765
                     /codon_start=3
                     /label=AUR1
                     /translation="RIDKLLGINMYTTAFSNSSVIFGAFPSLHSGCATMEALFFCYCFP
                     KLKPLFIAYVCWLWWSTMYLTHHYFVDLMAGSVLSYVIFQYTKYTHLPIVDTSLFCRWS
                     YTSIEKYDISKSDPLAADSNDIESVPLSNLELDFDLNMTDEPSVSPSLFDGSTSVSRSS
                     ATSITSLGVKRA"
     CDS             766..801
                     /codon_start=1
                     /label=linker
                     /translation="GGSGGSGGSGGS"
     CDS             802..1509
                     /codon_start=1
                     /label=yo-msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     3'UTR           1513..2002
                     /label=3' AUR1
     CDS             complement(2505..3308)
                     /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(3309..3524)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        3622..3726
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3727..4587
                     /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      4758..5346
                     /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 ccaagcttct
      241 agaattgata agctactcgg tattaatatg tatactacag ctttttcaaa ttcctccgtc
      301 attttcggtg cttttccttc actgcattcc gggtgtgcta ctatggaagc cctgtttttc
      361 tgttattgtt ttccaaaatt gaagcccttg tttattgctt atgtttgctg gttatggtgg
      421 tcaactatgt atctgacaca ccattatttt gtagacctta tggcaggttc tgtgctgtca
      481 tacgttattt tccagtacac aaagtacaca catttaccaa ttgtagatac atctcttttt
      541 tgcagatggt catacacttc aattgagaaa tacgatatat caaagagtga tccattggct
      601 gcagattcaa acgatatcga aagtgtccct ttgtccaact tggaacttga ctttgatctt
      661 aatatgactg atgaacccag tgtaagccct tcgttatttg atggatctac ttctgtttct
      721 cgttcgtccg ccacgtctat aacgtcacta ggtgtaaaga gggctggtgg cagtggcggt
      781 tctggaggct caggagggag catggatagc actgagagct tgttcacagg tgttgttcct
      841 atcttggtcg aattagacgg tgacgtaaac ggtcataagt tcagtgttag aggtgaaggt
      901 gaaggtgacg ctactaacgg taaattgaca ttgaagttta tttgtactac aggtaaattg
      961 ccagttcctt ggccaacttt ggttactaca ttgacatacg gtgttcaatg tttttcaaga
     1021 tacccagatc atatgaagca acatgatttc tttaagtctg ctatgccaga aggttacgtt
     1081 caagaaagaa ctatcacttt taaagatgat ggtacttaca agacaagagc agaggttaag
     1141 ttcgaaggtg acactttggt taacagaatc gaattgaagg gtatcgattt taaagaagat
     1201 ggtaatattt tgggtcataa attggaatac aactacaact cacataacgt ttacattaca
     1261 gctgataaac aaaagaatgg tattaaagca aacttcaaga tcagacataa tgttgaagat
     1321 ggttctgttc aattagcaga tcattaccaa caaaacactc caattggtga cggtccagtt
     1381 ttgttaccag ataaccatta cttgtctaca caatcaaaat tgtctaaaga tccaaacgaa
     1441 aagagagatc atatggtttt gttagaattt gttactgctg ctggtatcac aggtggcagt
     1501 ggcggttctt aatgagtatt ttatctgcaa ttacggatac ggttggtctt atgtagatac
     1561 atataaatat atatcttttt ctttcttttt cttagtcagg attgtcgttt agcataatat
     1621 acatgtagtt tatttaatca cataccactg attatcttta gaattttata aatttttgaa
     1681 ataaatgggt ggcttttaat ggtgtctatg ttaagtgagg cttttagaat gctcttcctg
     1741 ctttgtttat tatatgtgta tgaaagatat gtatgtattt acatgtgttt gtagcgtccc
     1801 cagtcaaaac ctgtgcgcta tacctaaatg gattgataat cttcattcac taattctaaa
     1861 atagacttct tccccaaaga acggtgtaac gatgaggctc tatccagctg cttatctaaa
     1921 tcaactttaa cgatggatga tcttatgaca cggggatctt tctttaaagt tcttagaatt
     1981 tcagactgta ccgcagctga tgggtaccga gctcgaattc actggccgtc gttttacaac
     2041 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2101 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2161 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2221 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     2281 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2341 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     2401 caccgaaacg cgcgagacga aagggcgggt aataactgat ataattaaat tgaagctcta
     2461 atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg
     2521 catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca
     2581 tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc
     2641 acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca
     2701 ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca
     2761 ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat
     2821 tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt
     2881 tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca
     2941 ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc
     3001 aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac
     3061 ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata
     3121 tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat
     3181 tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata
     3241 ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct
     3301 ttcgacatga tttatcttcg tttcctgcat gtttttgttc tgtgcagttg ggttaagaat
     3361 actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg
     3421 tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaaaga
     3481 aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaattgaaa agctggcctc
     3541 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     3601 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3661 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3721 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3781 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3841 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3901 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3961 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4021 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4081 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4141 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4201 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     4261 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     4321 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     4381 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     4441 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     4501 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4561 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     4621 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4681 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4741 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4801 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4861 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4921 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4981 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5041 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5101 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5161 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5221 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5281 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5341 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5401 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5461 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5521 gcggaaga
//
