LOCUS       Exported                7608 bp DNA     circular SYN 27-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7608)
  AUTHORS   Institute for Molecular Bioscience
  TITLE     Direct Submission
  JOURNAL   Exported Jan 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=Rseq_ScURA3t
     protein_bind    259..275
                     /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(283..313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    328..349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      637..1225
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1396..2256)
                     /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"
     promoter        complement(2257..2361)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2855..3027
                     /label=S. cerevisiae URA3 promoter
     gene            3028..3831
                     /product="Kluyveromyces lactis URA3"
                     /label=K. lactis URA3
     terminator      3832..3949
                     /label=K. lactis URA3 terminator
     terminator      3832..3949
                     /label=K. lactis URA3 terminator
     primer_bind     3884..3903
                     /label=Rseq_KlURA3t
     terminator      3961..4105
                     /label=HIS5 terminator
     primer_bind     4076..4128
                     /label=mRuby3_HIS5t_rv
     CDS             complement(4106..4819)
                     /codon_start=1
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     primer_bind     complement(4801..4846)
                     /label=mRuby3_AG4TGGA_fw
     primer_bind     4820..4876
                     /label=yEGFP_AG4TGGA_rv
     CDS             complement(4847..5572)
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae
                     and C. albicans"
                     /translation="MGSSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGI
                     KVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVL
                     LEFVTAAGITHGMDELYKRS"
     primer_bind     complement(5537..5582)
                     /label=yEGFP_BamHI_VP2C_fw
     gene            complement(5573..5770)
                     /label=VP2C
     promoter        5780..6447
                     /label=GAL10/GAL1 bi-directional promoter
                     /note="Obtained from plasmid pILGFPB5A"
     primer_bind     complement(5832..5862)
                     /label=Fseq_GAL10p
     protein_bind    5995..6112
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating
                     Gal4-dependent induction"
     primer_bind     6364..6395
                     /label=Fseq_GAL1p
     gene            6457..7599
                     /label=deltaVP1
     terminator      join(7608,1..234)
                     /label=S. cerevisiae URA3 terminator
ORIGIN
        1 ctagtaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
       61 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
      121 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
      181 cccgggaatc tcggtcgtaa tgatttaaat gcatgcaagc ttggcgtaat catggtcata
      241 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
      301 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
      361 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      421 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      481 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      541 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1501 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2341 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     2401 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     2461 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2521 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2581 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     2641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     2701 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2761 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2821 ccagtcacga cgttgtaaaa cgacggccag tgaattcatt taaatttttt tgattcggta
     2881 atctccgagc agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     2941 atatgtggtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     3001 aaaacctgca ggaaacgaag ataaaacatg tccacaaaat catataccag tagagctgag
     3061 actcatgcaa gtccggttgc atcgaaactt ttacgtttaa tggatgaaaa gaagaccaat
     3121 ttgtgtgctt ctcttgacgt tcgttcgact gatgagctat tgaaacttgt tgaaacgttg
     3181 ggtccataca tttgcctttt gaaaacacac gttgatatct tggatgattt cagttatgag
     3241 ggtactgtcg ttccattgaa agcattggca gagaaataca agttcttgat atttgaggac
     3301 agaaaattcg ccgatatcgg taacacagtc aaattacaat atacatcggg cgtttaccgt
     3361 atcgcagaat ggtctgatat caccaacgcc cacggggtta ctggtgctgg tattgttgct
     3421 ggcttgaaac aaggtgcgca agaggtcacc aaagaaccaa ggggattatt gatgcttgct
     3481 gaattgtctt ccaagggttc tctagcacac ggtgaatata ctaagggtac cgttgatatt
     3541 gcaaagagtg ataaagattt cgttattggg ttcattgctc agaacgatat gggaggaaga
     3601 gaagaagggt ttgattggct aatcatgacc ccaggtgtag gtttagacga caaaggcgat
     3661 gcattgggtc agcagtacag aaccgtcgac gaagttgtaa gtggtggatc agatatcatc
     3721 attgttggca gaggactttt cgccaagggt agagatccta aggttgaagg tgaaagatac
     3781 agaaatgctg gatgggaagc gtaccaaaag agaatcagcg ctccccatta attatacagg
     3841 aaacttaata gaacaaatca catatttaat ctaatagcca cctgcattgg cacggtgcaa
     3901 cactcacttc aacttcatct tacaaaagat cacgtgatct gttgtattgg gatctctaga
     3961 gtaacaatat catgagacct tttatagaag tggcgccaaa actaaatgta tttgaaaata
     4021 caaaaaacgc acaggtttct ttatatatgt atatatatat atatatatat ataaaactgt
     4081 acatatactg tttaaattaa tctatttact tgtacagctc gtccatgcca ccaccaaggt
     4141 tgctgtattt ggcaactgcc acttcgcgtt gcactacgta ggtttcattg tcactctcct
     4201 cgatcctttc caggcggtga tcaacggcat ggacaccggg catcttgatg ttcccgacgg
     4261 tcttttttga cctgtaagtt gtcacgaagt tgcagtgcag atggccacca ccatcaactt
     4321 tcagtgcgat gtcagtgtat cctctcagac caccatctgc tggatacatc atctctgtat
     4381 taggctccca acccttggtc ttcttctgca tcacgggacc attggaggga aagtttaccc
     4441 ctctgacctt gacgttgtag acgagctcgc catcctcaag gctggtgtcc tgcgtgacgg
     4501 tgacgactcc accatcttcg tatctcgtaa ctctttccca agtaaaaccc tcaggaaagg
     4561 actgtttaaa gaaatcaggg atgtcggccg ggtacttgat aaaagtacgg ctgccataca
     4621 tgaacgacgt ggcaagaatg tcaaaggcaa atggcagggg tcctccctcg atgactttga
     4681 tcctcatggt ttgcactccc tcgtacggtc tgccttctcc ttcacctgtg catttgaatt
     4741 ggtggccgtt gaccgaacct tccatgacca ccttcatacg catattttcc ttgatcagct
     4801 cttcgccctt agacaccata gcaccaccag tacctcctcc accagcagat cttttgtaca
     4861 attcatccat accatgggta ataccagcag cagtaacaaa ttctaacaag accatgtggt
     4921 ctctcttttc gtttggatct ttggataagg cagattgagt ggataagtaa tggttgtctg
     4981 gtaacaagac tggaccatca ccaattggag tattttgttg ataatggtca gctaattgaa
     5041 cagaaccatc ttcaatgttg tgtctaattt tgaagttaac tttgatacca ttcttttgtt
     5101 tgtcagccat gatgtaaaca ttgtgagagt tatagttgta ttccaatttg tgacctaaaa
     5161 tgttaccatc ttctttaaaa tcaatacctt ttaattcgat tctattaact aaggtatcac
     5221 cttcaaactt gacttcagct ctggtcttgt agttaccgtc atctttgaaa aaaatagttc
     5281 tttcttgaac ataaccttct ggcatggcag acttgaaaaa gtcatgttgt ttcatatgat
     5341 ctgggtatct cgcaaaacat tgaacaccat aaccgaaagt agtgactaag gttggccatg
     5401 gaactggcaa tttaccagta gtacaaataa attttaaggt caatttaccg taagtagcat
     5461 caccttcacc ttcaccggag acagaaaatt tgtgaccatt aacatcacca tctaattcaa
     5521 ccaaaattgg gacaacacca gtgaataatt cttcaccttt agaggatccc atagaaccgc
     5581 cgccaccgga gccaccgcca ccagaaccac caccacctgg agtgatgtca ccatacaaac
     5641 ccaaaatcaa tggcaacatc caatctggag taactctttg atgagaaaca acttgagctt
     5701 gataaaattt aataacttca ccagattcat cttgaatatg agcagctgga gttggaccac
     5761 cattacccat tttatcgatt tatattgaat tttcaaaaat tcttactttt tttttggatg
     5821 gacgcaaaga agtttaataa tcatattaca tggcattacc accatataca tatccatatc
     5881 taatcttact tatatgttgt ggaaatgtaa agagccccat tatcttagcc taaaaaaacc
     5941 ttctctttgg aactttcagt aatacgctta actgctcatt gctatattga agtacggatt
     6001 agaagccgcc gagcgggcga cagccctccg acggaagact ctcctccgtg cgtcctcgtc
     6061 ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa
     6121 agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc
     6181 cacaaacctt caaattaacg aatcaaatta acaaccatag gatgataatg cgattagttt
     6241 tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga
     6301 tatataaatg gaaaagctgc ataaccactt taactaatac tttcaacatt ttcagtttgt
     6361 attacttctt attcaaatgt cataaaagta tcaacaaaaa attgttaata tacctctata
     6421 ctttaacgtc aaggagaaaa aactatagct agcaaaatgg cttcaggtgt aagtaaatgc
     6481 gaaaccaaat gcacaaaggc ttgccctaga ccagctccag ttcctaagtt attgattaaa
     6541 ggtggtatgg aagttttgga tttggttaca ggtccagatt ctgttactga aatcgaagca
     6601 tttttgaacc caagaatggg tcaaccacca acaccagaat ctttaactga aggtggtcaa
     6661 tattacggtt ggtcaagagg tattaatttg gctacatctg atactgaaga ttcaccaggt
     6721 aataatacat taccaacttg gtctatggca aaattgcaat taccaatgtt gaacgaagat
     6781 ttgacttgtg atactttgca aatgtgggaa gcagtttcag ttaaaacaga agttgttggt
     6841 tctggttctt tgttggatgt tcatggtttt aataagccaa cagatactgt taacacaaag
     6901 ggtatttcta ctccagttga aggttcacaa tatcatgttt ttgctgttgg tggtgaacca
     6961 ttggatttgc aaggtttagt tacagatgca agaactaagt acaaggaaga aggtgttgtt
     7021 actattaaaa caatcactaa gaaagatatg gttaataagg atcaagtttt gaacccaatt
     7081 tcaaaggcta aattggataa ggatggcatg tacccagttg aaatttggca tccagatcca
     7141 gctaaaaatg aaaacacaag atacttcggt aattacactg gtggtactac aactccacca
     7201 gttttgcaat tcactaacac tttgacaact gttttgttag atgaaaatgg tgttggtcca
     7261 ttatgtaaag gtgaaggttt gtacttatct tgcgttgata ttatgggttg gagagttaca
     7321 agaaactacg atgttcatca ttggagaggt ttgccaagat acttcaagat tactttaaga
     7381 aaaagatggg ttaaaaatcc atacccaatg gcttctttga tttcttcttt gtttaataat
     7441 atgttaccac aagttcaagg tcaaccaatg gaaggtgaaa acacacaagt tgaagaagtt
     7501 agagtttacg atggtactga accagttcca ggtgaccctg acatgacaag atatgtagat
     7561 agattcggta aaactaaaac tgtattccct ggtaattaag cggccgca
//
