LOCUS       Exported                6991 bp ds-DNA     circular SYN 01-FEB-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   LC_Cheah
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Feb 1, 2021 from SnapGene 5.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6991
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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 
                     toinhibit 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 
                     presenceof 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
     promoter        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
     primer_bind     3884..3903
                     /label=Rseq_KlURA3t
     terminator      3961..4103
                     /label=HIS5 terminator
     misc_feature    4107..4217
                     /label=mODC(425-461)
                     /note="ref. Hoyt 2003 and Takeuchi 2008"
     CDS             complement(4218..4943)
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiaeand
                     C. albicans"
                     /translation="MGSSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGI
                     KVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVL
                     LEFVTAAGITHGMDELYKRS"
     gene            complement(4989..5141)
                     /label=VP2C
     promoter        5151..5818
                     /label=GAL10/GAL1 bi-directional promoter
                     /note="Obtained from plasmid pILGFPB5A"
     primer_bind     complement(5203..5233)
                     /label=Fseq_GAL10p
     protein_bind    5366..5483
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediatingGal4-dependent
                     induction"
     primer_bind     5735..5766
                     /label=Fseq_GAL1p
     gene            5828..6982
                     /label=wtVP1
     terminator      join(6991,1..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 aaaactgtac
     4081 atatactgtt taaattaatc tatttaaaca ttaattcttg cagatgcaca agcagctgga
     4141 tgcctatcca tacctgattc ttgtgcacat gacattggca aagttccatc atcttgctct
     4201 tcaacttcag gaggaaaaga tcttttgtac aattcatcca taccatgggt aataccagca
     4261 gcagtaacaa attctaacaa gaccatgtgg tctctctttt cgtttggatc tttggataag
     4321 gcagattgag tggataagta atggttgtct ggtaacaaga ctggaccatc accaattgga
     4381 gtattttgtt gataatggtc agctaattga acagaaccat cttcaatgtt gtgtctaatt
     4441 ttgaagttaa ctttgatacc attcttttgt ttgtcagcca tgatgtaaac attgtgagag
     4501 ttatagttgt attccaattt gtgacctaaa atgttaccat cttctttaaa atcaatacct
     4561 tttaattcga ttctattaac taaggtatca ccttcaaact tgacttcagc tctggtcttg
     4621 tagttaccgt catctttgaa aaaaatagtt ctttcttgaa cataaccttc tggcatggca
     4681 gacttgaaaa agtcatgttg tttcatatga tctgggtatc tcgcaaaaca ttgaacacca
     4741 taaccgaaag tagtgactaa ggttggccat ggaactggca atttaccagt agtacaaata
     4801 aattttaagg tcaatttacc gtaagtagca tcaccttcac cttcaccgga gacagaaaat
     4861 ttgtgaccat taacatcacc atctaattca accaaaattg ggacaacacc agtgaataat
     4921 tcttcacctt tagaggatcc catagaaccg ccgccaccgg agccaccgcc accagaacca
     4981 ccaccacctg gagtgatgtc accatacaaa cccaaaatca atggcaacat ccaatctgga
     5041 gtaactcttt gatgagaaac aacttgagct tgataaaatt taataacttc accagattca
     5101 tcttgaatat gagcagctgg agttggacca ccattaccca ttttatcgat ttatattgaa
     5161 ttttcaaaaa ttcttacttt ttttttggat ggacgcaaag aagtttaata atcatattac
     5221 atggcattac caccatatac atatccatat ctaatcttac ttatatgttg tggaaatgta
     5281 aagagcccca ttatcttagc ctaaaaaaac cttctctttg gaactttcag taatacgctt
     5341 aactgctcat tgctatattg aagtacggat tagaagccgc cgagcgggcg acagccctcc
     5401 gacggaagac tctcctccgt gcgtcctcgt cttcaccggt cgcgttcctg aaacgcagat
     5461 gtgcctcgcg ccgcactgct ccgaacaata aagattctac aatactagct tttatggtta
     5521 tgaagaggaa aaattggcag taacctggcc ccacaaacct tcaaattaac gaatcaaatt
     5581 aacaaccata ggatgataat gcgattagtt ttttagcctt atttctgggg taattaatca
     5641 gcgaagcgat gatttttgat ctattaacag atatataaat ggaaaagctg cataaccact
     5701 ttaactaata ctttcaacat tttcagtttg tattacttct tattcaaatg tcataaaagt
     5761 atcaacaaaa aattgttaat atacctctat actttaacgt caaggagaaa aaactatagc
     5821 tagcaaaatg gctccaaaga gaaagtcagg tgtaagtaaa tgcgaaacca aatgcacaaa
     5881 ggcttgccct agaccagctc cagttcctaa gttattgatt aaaggtggta tggaagtttt
     5941 ggatttggtt acaggtccag attctgttac tgaaatcgaa gcatttttga acccaagaat
     6001 gggtcaacca ccaacaccag aatctttaac tgaaggtggt caatattacg gttggtcaag
     6061 aggtattaat ttggctacat ctgatactga agattcacca ggtaataata cattaccaac
     6121 ttggtctatg gcaaaattgc aattaccaat gttgaacgaa gatttgactt gtgatacttt
     6181 gcaaatgtgg gaagcagttt cagttaaaac agaagttgtt ggttctggtt ctttgttgga
     6241 tgttcatggt tttaataagc caacagatac tgttaacaca aagggtattt ctactccagt
     6301 tgaaggttca caatatcatg tttttgctgt tggtggtgaa ccattggatt tgcaaggttt
     6361 agttacagat gcaagaacta agtacaagga agaaggtgtt gttactatta aaacaatcac
     6421 taagaaagat atggttaata aggatcaagt tttgaaccca atttcaaagg ctaaattgga
     6481 taaggatggc atgtacccag ttgaaatttg gcatccagat ccagctaaaa atgaaaacac
     6541 aagatacttc ggtaattaca ctggtggtac tacaactcca ccagttttgc aattcactaa
     6601 cactttgaca actgttttgt tagatgaaaa tggtgttggt ccattatgta aaggtgaagg
     6661 tttgtactta tcttgcgttg atattatggg ttggagagtt acaagaaact acgatgttca
     6721 tcattggaga ggtttgccaa gatacttcaa gattacttta agaaaaagat gggttaaaaa
     6781 tccataccca atggcttctt tgatttcttc tttgtttaat aatatgttac cacaagttca
     6841 aggtcaacca atggaaggtg aaaacacaca agttgaagaa gttagagttt acgatggtac
     6901 tgaaccagtt ccaggtgacc ctgacatgac aagatatgta gatagattcg gtaaaactaa
     6961 aactgtattc cctggtaatt aagcggccgc a
//
