LOCUS       Exported                7633 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 7633)
  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..7633
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        2..669
                     /label=GAL10/GAL1 bi-directional promoter
                     /note="Obtained from plasmid pILGFPB5A"
     primer_bind     complement(54..85)
                     /label=Fseq_GAL1p
     protein_bind    337..454
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating
                     Gal4-dependent induction"
     primer_bind     587..617
                     /label=Fseq_GAL10p
     primer_bind     640..699
                     /label=ERG20_pGAL10_fw
     CDS             679..1734
                     /codon_start=1
                     /label=wtERG20
                     /translation="MASEKEIRRERFLNVFPKLVEELNASLLAYGMPKEACDWYAHSLN
                     YNTPGGKLNRGLSVVDTYAILSNKTVEQLGQEEYEKVAILGWCIELLQAYFLVADDMMD
                     KSITRRGQPCWYKVPEVGEIAINDAFMLEAAIYKLLKSHFRNEKYYIDITELFHEVTFQ
                     TELGQLMDLITAPEDKVDLSKFSLKKHSFIVTFKTAYYSFYLPVALAMYVAGITDEKDL
                     KQARDVLIPLGEYFQIQDDYLDCFGTPEQIGKIGTDIQDNKCSWVINKALELASAEQRK
                     TLDENYGKKDSVAEAKCKKIFNDLKIDQLYHEYEESVAKDLKAKISQVDESRGFKADVL
                     TAFLNKVYKRSK"
     primer_bind     complement(741..759)
                     /label=Fseq_ERG20
     misc_feature    1623..2131
                     /label=gBlock for AG4TGGAx2
     primer_bind     1628..1649
                     /label=Rseq_ERG20
     CDS             1801..3522
                     /codon_start=1
                     /product="Actinidia chinensis nerolidol/linalool synthase
                     Codon optimized for expression in Saccharomyces cerevisae "
                     /label=AcNES1
                     /translation="MATAAGPIATNNSPQNSNAYRTPIAPSVPITHKWSIAEDLTCISN
                     PSKHNNPQTGYRSFSDELYVKYEEKLEDVRKALREVEENPLEGLVMIDALQRLGIDYHF
                     RGEIGAFLQKQQIISSTPDGYPEHGLYEVSTLFRFLRQEGHNVTADVFNNFKDKEGRFR
                     SELSTDIRGLMSLYEASQLRIEGEDILDQAADFSSQLLGRWTKDPNHHEARLVSNTLTH
                     PYHKSLATFMGQKLSYMNCKGPNWDGVDNLQELAKMDLTIVQSIHQKEVFQVSQWWKDT
                     GLANELKLARNQPLKWYMWPMAALTDPRFSEERVELTKPISFIYIIDDIFDVYGTIEEL
                     TLFTDAVNRWELSAVEQLPDYMKVCFKALYDVTNEIAYKIYKKHGQNPIDSLQKTWASL
                     CNAFLVEAKWFASGHLPNAEEYLKNGIISSGVHVVLAHMFFLLGDGITQESVDLVDDYP
                     GISTSIATILRLSDDLGSAKDEDQDGYDGSYIECYMKEHKGSSVDSAREEVIRMISEAW
                     KCLNKECLSPNPFSESFRIGSLNMARMIPMMYSYDDNHNLPILEEHMKAMIYNTSL"
     primer_bind     complement(3500..3559)
                     /label=AcNES1_HIS5t_rv
     terminator      3529..3673
                     /label=HIS5 terminator
     terminator      3685..3802
                     /label=K. lactis URA3 terminator
     terminator      3685..3802
                     /label=K. lactis URA3 terminator
     primer_bind     complement(3731..3750)
                     /label=Rseq_KlURA3t
     gene            complement(3803..4606)
                     /product="Kluyveromyces lactis URA3"
                     /label=K. lactis URA3
     misc_feature    4607..4779
                     /label=S. cerevisiae URA3 promoter
     primer_bind     complement(4783..4799)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5273..5377
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5378..6238
                     /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      6409..6997
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7285..7306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7321..7351
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7359..7375
                     /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)."
     primer_bind     7383..7399
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      join(7400..7633,1)
                     /label=S. cerevisiae URA3 terminator
     primer_bind     7565..7584
                     /label=Rseq_ScURA3t
     primer_bind     complement(join(7612..7633,1..28))
                     /label=GAL1p_patch
ORIGIN
        1 ttatagtttt ttctccttga cgttaaagta tagaggtata ttaacaattt tttgttgata
       61 cttttatgac atttgaataa gaagtaatac aaactgaaaa tgttgaaagt attagttaaa
      121 gtggttatgc agcttttcca tttatatatc tgttaataga tcaaaaatca tcgcttcgct
      181 gattaattac cccagaaata aggctaaaaa actaatcgca ttatcatcct atggttgtta
      241 atttgattcg ttaatttgaa ggtttgtggg gccaggttac tgccaatttt tcctcttcat
      301 aaccataaaa gctagtattg tagaatcttt attgttcgga gcagtgcggc gcgaggcaca
      361 tctgcgtttc aggaacgcga ccggtgaaga cgaggacgca cggaggagag tcttccgtcg
      421 gagggctgtc gcccgctcgg cggcttctaa tccgtacttc aatatagcaa tgagcagtta
      481 agcgtattac tgaaagttcc aaagagaagg tttttttagg ctaagataat ggggctcttt
      541 acatttccac aacatataag taagattaga tatggatatg tatatggtgg taatgccatg
      601 taatatgatt attaaacttc tttgcgtcca tccaaaaaaa aagtaagaat ttttgaaaat
      661 tcaatataag gatccaaaat ggcttcagaa aaagaaatta ggagagagag attcttgaac
      721 gttttcccta aattagtaga ggaattgaac gcatcgcttt tggcttacgg tatgcctaag
      781 gaagcatgtg actggtatgc ccactcattg aactacaaca ctccaggcgg taagttaaat
      841 agaggtttgt ccgttgtgga cacgtatgct attctctcca acaagaccgt tgaacaattg
      901 gggcaagaag aatacgaaaa ggttgctatt ctaggttggt gcattgagtt gttgcaggct
      961 tacttcttgg tcgccgatga tatgatggac aagtccatta ccagaagagg ccaaccatgt
     1021 tggtacaagg ttcctgaagt tggggaaatt gccatcaatg acgcattcat gttagaggct
     1081 gctatctaca agcttttgaa atctcacttc agaaacgaaa aatactacat agatatcacc
     1141 gaattgttcc atgaagtcac cttccaaacc gaattgggcc aattgatgga cttaatcact
     1201 gcacctgaag acaaagtcga cttgagtaag ttctccctaa agaagcactc cttcatagtt
     1261 actttcaaga ctgcttacta ttctttctac ttgcctgtcg cattggctat gtacgttgcc
     1321 ggtatcacag atgaaaagga tttgaaacaa gccagagatg tcttgattcc attgggtgaa
     1381 tatttccaaa ttcaagatga ctacttagac tgcttcggta ccccagaaca gatcggtaag
     1441 atcggtacag atatccaaga taacaaatgt tcttgggtaa tcaacaaggc attagaactt
     1501 gcttccgcag aacaaagaaa gactttagac gaaaattacg gtaagaagga ctcagtcgca
     1561 gaagccaaat gcaaaaagat tttcaatgac ttgaaaatcg accagttata ccacgaatat
     1621 gaagagtctg ttgccaagga tttgaaggcc aagatctccc aagtcgacga gtctcgtggc
     1681 ttcaaagccg acgtcttaac tgcgtttttg aacaaagttt acaagagaag caaaagatct
     1741 gctggtggag gaggtactgg tggtgctgca ggaggcggag gtacaggagg tgcagagctc
     1801 atggctaccg cagcaggtcc tatcgcaact aacaactccc cacaaaactc caacgcttac
     1861 agaactccaa tcgctccttc cgtaccaatt actcataaat ggtctatagc tgaagatttg
     1921 acatgtattt ccaatcctag taagcacaat aaccctcaaa ctggttacag atcattttct
     1981 gacgaattat acgttaagta cgaagaaaag ttggaagatg ttagaaaagc attaagagaa
     2041 gttgaagaaa accctttgga aggtttagtt atgatagacg ctttgcaaag attgggtatc
     2101 gattaccatt tcagaggtga aattggtgca ttcttgcaaa agcaacaaat catatcttca
     2161 actccagatg gttaccctga acatggtttg tacgaagttt caacattgtt tagattctta
     2221 agacaagaag gtcacaatgt taccgctgac gtctttaata acttcaagga taaggaaggt
     2281 agattcagat cagaattgtc aacagatatt agaggtttga tgtccttata cgaagcaagt
     2341 caattgagaa tagaaggtga agacatctta gatcaagctg ctgatttctc cagtcaattg
     2401 ttaggtagat ggacaaaaga tcctaatcat cacgaagcca gattggtttc taacacttta
     2461 acacatccat accacaagtc attggctacc tttatgggtc aaaaattgtc ctacatgaac
     2521 tgcaagggtc caaactggga cggtgtcgat aatttgcaag aattagctaa gatggatttg
     2581 actatcgtac aaagtatcca tcaaaaggaa gtattccaag tttctcaatg gtggaaggat
     2641 acaggtttag ccaatgaatt gaaattggct agaaaccaac cattgaagtg gtatatgtgg
     2701 cctatggccg ctttaaccga tccaagattc agtgaagaaa gagttgaatt aactaagcct
     2761 atttctttta tatacatcat agatgacatc ttcgacgtct atggtaccat tgaagaattg
     2821 accttgttta ctgatgcagt taatagatgg gaattgtctg ccgtcgaaca attaccagac
     2881 tacatgaaag tatgtttcaa ggctttgtac gatgttacca acgaaatcgc atacaaaatc
     2941 tataaaaagc atggtcaaaa ccctattgat tccttgcaaa agacttgggc tagtttgtgc
     3001 aatgcatttt tagttgaagc aaagtggttc gcctctggtc acttgccaaa tgcagaagaa
     3061 tacttaaaga acggtatcat ctcttcaggt gttcatgttg tcttggccca catgtttttc
     3121 ttgttaggtg acggtattac acaagaatca gttgatttgg tagatgacta tccaggtatt
     3181 tccacaagta tcgcaaccat tttgagatta tctgatgact tgggttcagc caaagatgaa
     3241 gaccaagatg gttatgatgg ttcttacatc gaatgttaca tgaaggaaca taagggttcc
     3301 agtgtcgatt cagccagaga agaagtaata agaatgatct ccgaagcatg gaaatgtttg
     3361 aataaggaat gcttatcacc aaaccctttt tctgaatcat tcagaatagg ttccttgaat
     3421 atggctagaa tgatccctat gatgtactct tacgatgaca accataactt gccaatttta
     3481 gaagaacaca tgaaggcaat gatctataac acatctttat aactcgagat agattaattt
     3541 aaacagtata tgtacagttt tatatatata tatatatata tatacatata taaagaaacc
     3601 tgtgcgtttt ttgtattttc aaatacattt agttttggcg ccacttctat aaaaggtctc
     3661 atgatattgt tactctagag atcccaatac aacagatcac gtgatctttt gtaagatgaa
     3721 gttgaagtga gtgttgcacc gtgccaatgc aggtggctat tagattaaat atgtgatttg
     3781 ttctattaag tttcctgtat aattaatggg gagcgctgat tctcttttgg tacgcttccc
     3841 atccagcatt tctgtatctt tcaccttcaa ccttaggatc tctacccttg gcgaaaagtc
     3901 ctctgccaac aatgatgata tctgatccac cacttacaac ttcgtcgacg gttctgtact
     3961 gctgacccaa tgcatcgcct ttgtcgtcta aacctacacc tggggtcatg attagccaat
     4021 caaacccttc ttctcttcct cccatatcgt tctgagcaat gaacccaata acgaaatctt
     4081 tatcactctt tgcaatatca acggtaccct tagtatattc accgtgtgct agagaaccct
     4141 tggaagacaa ttcagcaagc atcaataatc cccttggttc tttggtgacc tcttgcgcac
     4201 cttgtttcaa gccagcaaca ataccagcac cagtaacccc gtgggcgttg gtgatatcag
     4261 accattctgc gatacggtaa acgcccgatg tatattgtaa tttgactgtg ttaccgatat
     4321 cggcgaattt tctgtcctca aatatcaaga acttgtattt ctctgccaat gctttcaatg
     4381 gaacgacagt accctcataa ctgaaatcat ccaagatatc aacgtgtgtt ttcaaaaggc
     4441 aaatgtatgg acccaacgtt tcaacaagtt tcaatagctc atcagtcgaa cgaacgtcaa
     4501 gagaagcaca caaattggtc ttcttttcat ccattaaacg taaaagtttc gatgcaaccg
     4561 gacttgcatg agtctcagct ctactggtat atgattttgt ggacatgttt tatcttcgtt
     4621 tcctgcaggt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     4681 caacaccaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     4741 tctgctcgga gattaccgaa tcaaaaaaat ttaaatgaat tcactggccg tcgttttaca
     4801 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     4861 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4921 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4981 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5041 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5101 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5161 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     5221 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5281 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5341 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5401 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5461 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5521 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5581 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5641 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5701 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5761 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5821 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5881 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5941 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6001 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6061 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6121 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6181 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6241 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6301 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6361 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6421 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6481 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6541 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6601 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6661 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6721 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6781 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6841 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6901 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6961 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7021 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7081 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     7141 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     7201 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     7261 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     7321 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7381 cacaggaaac agctatgacc atgattacgc caagcttgca tgcatttaaa tcattacgac
     7441 cgagattccc gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta
     7501 tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg
     7561 cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa
     7621 atagtcctac tag
//
