LOCUS       Exported                7556 bp DNA     circular SYN 29-NOV-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7556)
  AUTHORS   Jason Yu
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7556
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     primer_bind     164..182
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(220..242)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        566..973
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             974..2068
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(2740..2761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2952..2971
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3188..3210
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3202..3219
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3203..3219
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3226..3245
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3226..3244
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3247..3270
                     /label=p16NotI_GAP_f
     misc_feature    3253..3334
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3313..3365
                     /label=p16_GAPprom_f
     primer_bind     complement(3313..3365)
                     /label=GAPprom_p16_r
     primer_bind     complement(3313..3339)
                     /label=uni_p16_r
     primer_bind     3339..3365
                     /label=p16NotI_GAP_f
     promoter        3340..4006
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     primer_bind     3340..3361
                     /label=NotI_GAPpro_f
                     /note="cloning primer to create insert from the eYp plasmid
                     fluorophores"
     primer_bind     3850..3869
                     /label=GPD fw 1
     primer_bind     3897..3916
                     /label=GPD fw 2
     primer_bind     3924..3947
                     /label=p426-sense(GPD_pr)
     primer_bind     complement(3982..4012)
                     /label=mCherry_GAPprom_r
     primer_bind     complement(4007..4019)
                     /label=XhoI_GAPprom_r
     CDS             4031..4747
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yomSapphire
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     4760..4788
                     /label=mCherry_ADH1term_f
     terminator      4768..4955
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     4768..4783
                     /label=NheI_ADHIterm_f
     primer_bind     complement(4931..4955)
                     /label=p16SalI_ADH1_r
     primer_bind     complement(4939..4984)
                     /label=p16_ADH1term_r
     primer_bind     complement(4942..4955)
                     /label=SalI_ADH1term_r
                     /note="cloning primer to create insert from the eYp plasmid
                     fluorophores"
     primer_bind     4958..4984
                     /label=uni_p16_f
     misc_feature    4958..4962
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     promoter        complement(4974..4992)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4974..4992)
                     /label=T3
                     /note="T3 promoter, forward primer"
     primer_bind     complement(5013..5029)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5013..5029)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5026..5048)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5037..5053
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5061..5091)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5106..5127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5244..5261)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5415..6004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5495..5514)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6175..7035)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6798..6817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7036..7140)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(7170..7556,1..117)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt
       61 taatttatat atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa
      121 aaggacctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      181 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
      241 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
      301 gtcagcgggt gttggcggcg cgccatcgag gagaacttct agtatatcta catacctaat
      361 attattgcct tattaaaaat ggaatcccaa caattacatc aaaatccaca ttctcttcaa
      421 aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat ttataggata
      481 attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta aggcgcctga
      541 ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt aagatgcaag
      601 agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc
      661 gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct
      721 gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga
      781 accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat cacaatactt
      841 gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt agcaatcgtc
      901 ttactttcta acttttctta ccttttacat ttcagcaata tatatatata tatttcaagg
      961 atataccatt ctaatgtctg cccctaagaa gatcgtcgtt ttgccaggtg accacgttgg
     1021 tcaagaaatc acagccgaag ccattaaggt tcttaaagct atttctgatg ttcgttccaa
     1081 tgtcaagttc gatttcgaaa atcatttaat tggtggtgct gctatcgatg ctacaggtgt
     1141 tccacttcca gatgaggcgc tggaagcctc caagaaggct gatgccgttt tgttaggtgc
     1201 tgtgggtggt cctaaatggg gtaccggtag tgttagacct gaacaaggtt tactaaaaat
     1261 ccgtaaagaa cttcaattgt acgccaactt aagaccatgt aactttgcat ccgactctct
     1321 tttagactta tctccaatca agccacaatt tgctaaaggt actgacttcg ttgttgtcag
     1381 agaattagtg ggaggtattt actttggtaa gagaaaggaa gacgatggtg atggtgtcgc
     1441 ttgggatagt gaacaataca ccgttccaga agtgcaaaga atcacaagaa tggccgcttt
     1501 catggcccta caacatgagc caccattgcc tatttggtcc ttggataaag ctaatgtttt
     1561 ggcctcttca agattatgga gaaaaactgt ggaggaaacc atcaagaacg aattccctac
     1621 attgaaggtt caacatcaat tgattgattc tgccgccatg atcctagtta agaacccaac
     1681 ccacctaaat ggtattataa tcaccagcaa catgtttggt gatatcatct ccgatgaagc
     1741 ctccgttatc ccaggttcct tgggtttgtt gccatctgcg tccttggcct ctttgccaga
     1801 caagaacacc gcatttggtt tgtacgaacc atgccacggt tctgctccag atttgccaaa
     1861 gaataaggtc aaccctatcg ccactatctt gtctgctgca atgatgttga aattgtcatt
     1921 gaacttgcct gaagaaggta aggccattga agatgcagtt aaaaaggttt tggatgcagg
     1981 tatcagaact ggtgatttag gtggttccaa cagtaccacc gaagtcggtg atgctgtcgc
     2041 cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt tttttatgat atttgtacat
     2101 aaactttata aatgaaattc ataatagaaa cgacacgaaa ttacaaaatg gaatatgttc
     2161 atagggtaga cgaaactata tacgcaatct acatacattt atcaagaagg agaaaaagga
     2221 ggatgtaaag gaatacaggt aagcaaattg atactaatgg ctcaacgtga taaggaaaaa
     2281 gaattgcact ttaacattaa tattgacaag gaggagggca ccacacaaaa agttaggtgt
     2341 aacagaaaat catgaaacta tgattcctaa tttatatatt ggaggatttt ctctaaaaaa
     2401 aaaaaaatac aacaaataaa aaacactcaa tgacctgacc atttgatgga gtttaagtca
     2461 ataccttctt gaaccatttc ccataatggt gaaagttccc tcaagaattt tactctgtca
     2521 gaaacggcct tacgacgtag tcgaggatcc tatgcggtgt gaaataccgc acagatgcgt
     2581 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     2641 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2701 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2761 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2821 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2881 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2941 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc tggcaagtgt
     3001 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3061 gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     3121 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     3181 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     3241 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg gtcccccggg
     3301 ctgcaggaat tcgatatcaa gcttatcgat accgtcgact cattatcaat actgccattt
     3361 caaagaatac gtaaataatt aatagtagtg attttcctaa ctttatttag tcaaaaaatt
     3421 agccttttaa ttctgctgta acccgtacat gcccaaaata gggggcgggt tacacagaat
     3481 atataacatc gtaggtgtct gggtgaacag tttattcctg gcatccacta aatataatgg
     3541 agcccgcttt ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa aatattgttt
     3601 tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga gaacaggggc
     3661 acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc tggagtaaat
     3721 gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca ccttctatta
     3781 ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca gttccctgaa
     3841 attattcccc tacttgacta ataagtatat aaagacggta ggtattgatt gtaattctgt
     3901 aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt tttagtttta
     3961 aaacaccaag aacttagttt cgaataaaca cacataaaca aacaaaggtg acggtgctgg
     4021 tttaattaac atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
     4081 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
     4141 tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg
     4201 gccaacctta gtcactactt tttcttatgg tgttcaatgt tttgctagat acccagatca
     4261 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     4321 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     4381 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     4441 aggtcacaaa ttggaataca actttaactc tcacaatgtt tacatcatgg ctgacaaaca
     4501 aaagaatggt atcaaagtta acttcaaaat tagacacaac attgaagatg gttctgttca
     4561 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     4621 caaccattac ttatccattc aatctgcctt atccaaagat ccaaacgaaa agagagacca
     4681 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     4741 caaataaggc gcgccacttc taaataagcg aatttcttat gatttatgat ttttattatt
     4801 aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa
     4861 cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag
     4921 tatgaggtcg ctcttattga ccacacctct accggcagta ccagcttttg ttccctttag
     4981 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5041 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5101 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5161 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5221 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5281 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5341 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5401 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5461 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tcccccctgg
     5521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5821 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6241 ctgactgccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgaaaaa aagcggttag
     6601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7141 cacatttccc cgaaaagtgc cacctgacgg atcgcttgcc tgtaacttac acgcgcctcg
     7201 tatcttttaa tgatggaata atttgggaat ttactctgta tttatttatt tttatgtttt
     7261 gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa
     7321 aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga
     7381 caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac
     7441 aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg
     7501 agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag agtctt
//
