LOCUS       Exported                6972 bp DNA     circular SYN 08-SEP-2021
DEFINITION  Plasmid for chromosomal tagging of Sec2 with three copies of msGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
COMMENT     Linearize with MfeI for pop-in integration at SEC2, then select for 
            pop-outs on 5-FOA.
FEATURES             Location/Qualifiers
     source          1..6972
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     CDS             250..943
                     /codon_start=2
                     /label=SEC2
                     /translation="DKALWKQDAKRPFSSSSAEESQKSDAFDFESGDMENEITGESSSD
                     ESSSDGSSTDNSTADSSSEDESSLADSTTSSADSSSPESIDNGEGDDTVTKDDKSSIKS
                     ANNNEENSDCGDKKGRSIIKKKAPQRKIQKKKLLQDLDDLEEQFREESAIDQTEFENAE
                     SNVKQNISSKRASSGDENSKKDNNEKTLKTNLTIGDKTQEQIGENSPSSGLHASSSNDD
                     NFDDAQEQQ"
     CDS             944..988
                     /codon_start=1
                     /label=Linker
                     /translation="TDPGSSGSSGSSGAS"
     CDS             989..1703
                     /codon_start=1
                     /label=msGFP
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGSSGSSG"
     CDS             1709..2423
                     /codon_start=1
                     /label=msGFP
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGSSGSSG"
     CDS             2429..3145
                     /codon_start=1
                     /label=msGFP
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGSSGSSG"
     3'UTR           3155..3359
                     /label=3' SEC2
     CDS             complement(3949..4752)
                     /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(4753..4973)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        5066..5170
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5171..6031
                     /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      6202..6790
                     /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 atgcctgcag gacaaagctc tttggaagca ggacgcaaaa agaccttttt cttcatctag
      301 cgcagaagaa agtcaaaaga gcgacgcttt tgactttgaa agtggagata tggaaaatga
      361 aattacgggt gaatcttcaa gtgatgaatc ctcttcagat gggtcttcca cagataattc
      421 aaccgcagat agttcatcag aagatgaatc ttctctagca gattctacta caagtagcgc
      481 agattctagt tcaccagaga gtatagacaa cggtgaaggt gatgacacgg taacgaaaga
      541 tgacaaaagt agcattaagt cggcaaataa taatgaagaa aacagcgatt gcggagataa
      601 aaaaggtcgt tctataataa agaaaaaggc tcctcaacgc aagatacaaa agaagaagtt
      661 actccaagat ttggatgatt tagaagagca atttagagaa gaaagcgcaa ttgatcagac
      721 tgaatttgaa aacgctgagt ctaatgtaaa acaaaatata tcctcgaaga gggcatcttc
      781 tggagacgag aactctaaga aggataataa tgaaaaaact ttaaagacaa acttgacgat
      841 aggcgacaaa acccaagaac aaatagggga aaatagtcct agtagcggat tgcatgcgtc
      901 ttcaagcaat gatgacaatt tcgatgatgc ccaggaacag caaacggatc caggaagttc
      961 ggggtccagc ggttcttcag gcgctagcat ggatagcact gagagcctgt tcaccggggt
     1021 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg
     1081 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     1141 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1201 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1261 ctacgtccag gagcgcacca tcaccttcaa ggacgacggc acctacaaga cccgcgccga
     1321 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1381 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta
     1441 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
     1501 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1561 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     1621 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
     1681 cggcagctcc ggctccagcg ggtctagcat ggatagcact gagagcctgt tcaccggggt
     1741 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg
     1801 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     1861 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1921 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1981 ctacgtccag gagcgcacca tcaccttcaa ggacgacggc acctacaaga cccgcgccga
     2041 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2101 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta
     2161 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
     2221 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2281 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     2341 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
     2401 cggcagctcc ggctccagcg ggtctagcat ggatagcact gagagcctgt tcaccggggt
     2461 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgcgcgg
     2521 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg
     2581 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     2641 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2701 ctacgtccag gagcgcacca tcaccttcaa ggacgacggc acctacaaga cccgcgccga
     2761 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2821 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaacgtcta
     2881 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
     2941 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3001 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     3061 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactca
     3121 cggcagctcc ggctccagcg ggtaaagcgg ccgcattata taatatttaa ttttatgtgt
     3181 cctaatggac atttgagttc cttaagatgt tcaacaactt agaaaaatat aggtaatgat
     3241 aaagaatgaa tcgcatgtat tgttttaata ttatgtcaat actttgaact taaaaacaga
     3301 gtaaagaagt ggttctaaaa ttacacgtat taaagggatt aattactaca tattcattct
     3361 caatcatcac tatcaccttg gctcaaaaca actctagact ttttgccaaa aaggcggatg
     3421 gcctcttcat cttcatcgct ttcatcggta ccgagctcga attcactggc cgtcgtttta
     3481 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3541 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3601 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3661 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3721 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3781 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3841 tcatcaccga aacgcgcgag acgaaagggc gggtaataac tgatataatt aaattgaagc
     3901 tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg
     3961 gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt
     4021 agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga
     4081 gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc
     4141 cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg
     4201 agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt
     4261 atattctcca gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct
     4321 aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac
     4381 cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta
     4441 ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt
     4501 gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa
     4561 gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag
     4621 taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat
     4681 gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt
     4741 agctttcgac atgatttatc ttcgtttcct gcatgttttt gttctgtgca gttgggttaa
     4801 gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag
     4861 tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca
     4921 aagaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctgg
     4981 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5041 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5101 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5161 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5221 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5281 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5341 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5401 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5461 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5521 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5581 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5641 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5701 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5761 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5821 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5881 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5941 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6001 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6061 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6121 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6181 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6241 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6301 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6361 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6421 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6481 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6541 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6601 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6661 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6721 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6781 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6841 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6901 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6961 ggaagcggaa ga
//
