LOCUS       Exported                6790 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 6790)
  AUTHORS   Institute for Molecular Bioscience
  TITLE     Direct Submission
  JOURNAL   Exported Jan 27, 2022 from SnapGene 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6790
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..32
                     /label=Fseq_GAL1p
     primer_bind     58..110
                     /label=VP1_NheI_GAL1p_fw
     gene            94..1044
                     /label=deltaVP1 (C-ter 63 aa deleted)
     primer_bind     complement(1025..1077)
                     /label=VP1dC63_URA3t_rv
     misc_feature    1034..1044
                     /label=deltaC63 truncation site
     terminator      1056..1290
                     /label=S. cerevisiae URA3 terminator
     primer_bind     complement(1106..1125)
                     /label=Rseq_ScURA3t
     primer_bind     complement(1291..1307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1315..1331
                     /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(1339..1369)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1384..1405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1693..2281
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2452..3312)
                     /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(3313..3417)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3891..3907
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3911..4083
                     /label=S. cerevisiae URA3 promoter
     gene            4084..4887
                     /product="Kluyveromyces lactis URA3"
                     /label=K. lactis URA3
     terminator      4888..5005
                     /label=K. lactis URA3 terminator
     primer_bind     4940..4959
                     /label=Rseq_KlURA3t
     primer_bind     4991..5043
                     /label=HIS5t_KlURA3t_rv
     terminator      5017..5159
                     /label=HIS5 terminator
     misc_feature    5163..5273
                     /label=mODC(425-461)
                     /note="ref. Hoyt 2003 and Takeuchi 2008"
     CDS             complement(5274..5999)
                     /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(5978..6020)
                     /label=yEGFP_VP2_fw
     primer_bind     5978..6020
                     /label=VP2_yEGFP_rv
     gene            complement(6045..6197)
                     /label=VP2C
     primer_bind     complement(6177..6236)
                     /label=VP2_ClaI_GAL10p_fw
     promoter        join(6207..6790,1..84)
                     /label=GAL10/GAL1 bi-directional promoter
                     /note="Obtained from plasmid pILGFPB5A"
     primer_bind     complement(6259..6289)
                     /label=Fseq_GAL10p
     protein_bind    6422..6539
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
ORIGIN
        1 acttcttatt caaatgtcat aaaagtatca acaaaaaatt gttaatatac ctctatactt
       61 taacgtcaag gagaaaaaac tatagctagc aaaatggctt caggtgtaag taaatgcgaa
      121 accaaatgca caaaggcttg ccctagacca gctccagttc ctaagttatt gattaaaggt
      181 ggtatggaag ttttggattt ggttacaggt ccagattctg ttactgaaat cgaagcattt
      241 ttgaacccaa gaatgggtca accaccaaca ccagaatctt taactgaagg tggtcaatat
      301 tacggttggt caagaggtat taatttggct acatctgata ctgaagattc accaggtaat
      361 aatacattac caacttggtc tatggcaaaa ttgcaattac caatgttgaa cgaagatttg
      421 acttgtgata ctttgcaaat gtgggaagca gtttcagtta aaacagaagt tgttggttct
      481 ggttctttgt tggatgttca tggttttaat aagccaacag atactgttaa cacaaagggt
      541 atttctactc cagttgaagg ttcacaatat catgtttttg ctgttggtgg tgaaccattg
      601 gatttgcaag gtttagttac agatgcaaga actaagtaca aggaagaagg tgttgttact
      661 attaaaacaa tcactaagaa agatatggtt aataaggatc aagttttgaa cccaatttca
      721 aaggctaaat tggataagga tggcatgtac ccagttgaaa tttggcatcc agatccagct
      781 aaaaatgaaa acacaagata cttcggtaat tacactggtg gtactacaac tccaccagtt
      841 ttgcaattca ctaacacttt gacaactgtt ttgttagatg aaaatggtgt tggtccatta
      901 tgtaaaggtg aaggtttgta cttatcttgc gttgatatta tgggttggag agttacaaga
      961 aactacgatg ttcatcattg gagaggtttg ccaagatact tcaagattac tttaagaaaa
     1021 agatgggtta aaaatccata cccataagcg gccgcactag taggactatt tgcaaaggga
     1081 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     1141 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     1201 caaattagag cttcaattta attatatcag ttattacccg ggaatctcgg tcgtaatgat
     1261 ttaaatgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1321 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1381 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1441 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1501 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1561 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2101 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2401 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2461 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2521 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2581 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2641 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2701 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2761 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2821 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2881 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2941 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3001 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3061 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3121 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3181 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3241 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3301 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3361 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3421 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     3481 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     3541 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     3601 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     3661 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     3721 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     3781 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     3841 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     3901 ggccagtgaa ttcatttaaa tttttttgat tcggtaatct ccgagcagaa ggaagaacga
     3961 aggaaggagc acagacttag attggtatat atacgcatat gtggtgttga agaaacatga
     4021 aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa
     4081 aacatgtcca caaaatcata taccagtaga gctgagactc atgcaagtcc ggttgcatcg
     4141 aaacttttac gtttaatgga tgaaaagaag accaatttgt gtgcttctct tgacgttcgt
     4201 tcgactgatg agctattgaa acttgttgaa acgttgggtc catacatttg ccttttgaaa
     4261 acacacgttg atatcttgga tgatttcagt tatgagggta ctgtcgttcc attgaaagca
     4321 ttggcagaga aatacaagtt cttgatattt gaggacagaa aattcgccga tatcggtaac
     4381 acagtcaaat tacaatatac atcgggcgtt taccgtatcg cagaatggtc tgatatcacc
     4441 aacgcccacg gggttactgg tgctggtatt gttgctggct tgaaacaagg tgcgcaagag
     4501 gtcaccaaag aaccaagggg attattgatg cttgctgaat tgtcttccaa gggttctcta
     4561 gcacacggtg aatatactaa gggtaccgtt gatattgcaa agagtgataa agatttcgtt
     4621 attgggttca ttgctcagaa cgatatggga ggaagagaag aagggtttga ttggctaatc
     4681 atgaccccag gtgtaggttt agacgacaaa ggcgatgcat tgggtcagca gtacagaacc
     4741 gtcgacgaag ttgtaagtgg tggatcagat atcatcattg ttggcagagg acttttcgcc
     4801 aagggtagag atcctaaggt tgaaggtgaa agatacagaa atgctggatg ggaagcgtac
     4861 caaaagagaa tcagcgctcc ccattaatta tacaggaaac ttaatagaac aaatcacata
     4921 tttaatctaa tagccacctg cattggcacg gtgcaacact cacttcaact tcatcttaca
     4981 aaagatcacg tgatctgttg tattgggatc tctagagtaa caatatcatg agacctttta
     5041 tagaagtggc gccaaaacta aatgtatttg aaaatacaaa aaacgcacag gtttctttat
     5101 atatgtatat atatatatat atatataaaa ctgtacatat actgtttaaa ttaatctatt
     5161 taaacattaa ttcttgcaga tgcacaagca gctggatgcc tatccatacc tgattcttgt
     5221 gcacatgaca ttggcaaagt tccatcatct tgctcttcaa cttcaggagg aaaagatctt
     5281 ttgtacaatt catccatacc atgggtaata ccagcagcag taacaaattc taacaagacc
     5341 atgtggtctc tcttttcgtt tggatctttg gataaggcag attgagtgga taagtaatgg
     5401 ttgtctggta acaagactgg accatcacca attggagtat tttgttgata atggtcagct
     5461 aattgaacag aaccatcttc aatgttgtgt ctaattttga agttaacttt gataccattc
     5521 ttttgtttgt cagccatgat gtaaacattg tgagagttat agttgtattc caatttgtga
     5581 cctaaaatgt taccatcttc tttaaaatca atacctttta attcgattct attaactaag
     5641 gtatcacctt caaacttgac ttcagctctg gtcttgtagt taccgtcatc tttgaaaaaa
     5701 atagttcttt cttgaacata accttctggc atggcagact tgaaaaagtc atgttgtttc
     5761 atatgatctg ggtatctcgc aaaacattga acaccataac cgaaagtagt gactaaggtt
     5821 ggccatggaa ctggcaattt accagtagta caaataaatt ttaaggtcaa tttaccgtaa
     5881 gtagcatcac cttcaccttc accggagaca gaaaatttgt gaccattaac atcaccatct
     5941 aattcaacca aaattgggac aacaccagtg aataattctt cacctttaga ggatcccata
     6001 gaaccgccgc caccggagcc accgccacca gaaccaccac cacctggagt gatgtcacca
     6061 tacaaaccca aaatcaatgg caacatccaa tctggagtaa ctctttgatg agaaacaact
     6121 tgagcttgat aaaatttaat aacttcacca gattcatctt gaatatgagc agctggagtt
     6181 ggaccaccat tacccatttt atcgatttat attgaatttt caaaaattct tacttttttt
     6241 ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc atatacatat
     6301 ccatatctaa tcttacttat atgttgtgga aatgtaaaga gccccattat cttagcctaa
     6361 aaaaaccttc tctttggaac tttcagtaat acgcttaact gctcattgct atattgaagt
     6421 acggattaga agccgccgag cgggcgacag ccctccgacg gaagactctc ctccgtgcgt
     6481 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
     6541 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
     6601 ctggccccac aaaccttcaa attaacgaat caaattaaca accataggat gataatgcga
     6661 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
     6721 taacagatat ataaatggaa aagctgcata accactttaa ctaatacttt caacattttc
     6781 agtttgtatt
//
