LOCUS       Exported                6529 bp ds-DNA     circular SYN 16-DEC-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6529)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 16, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6529
                     /organism="synthetic DNA construct"
                     /label=/dnas_title=pCL10_features_adjusted
                     /note="/dnas_title=pCL10_features_adjusted"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /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      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2243..2261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2334..2957
                     /label=GAL1pr
                     /note="I tested YFP expression and it works"
     protein_bind    2516..2633
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     misc_feature    2516..2529
                     /label=GAL4 BS
     misc_feature    2535..2551
                     /label=GAL4 BS
     CDS             2967..4217
                     /codon_start=1
                     /label=yEVenus-CLN2PEST
                     /note="yEVenus-CLN2PEST"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYKASNLNISRKLTISTPSCSFENSNSTSIPSPASSSQSHTPMRNM
                     SSLSDNSVFSRNMEQSSPITPSMYQFGQQQSNSICGSTVSVNSLVNTNNKQRIYEQITG
                     PNSNNATNDYIDLLNLNESNKENQNPATAHYLNGGPPKTSFINHGMFPSPTGTINSGKS
                     SSASSLISFGMGNTQVI"
     misc_feature    2967..3680
                     /label=yEVenus from pKT0090 (addgene), but no stop codon
                     /note="yEVenus as pulled from pKT0090 plasmid addgene, just
                     missing stop codon"
     CDS             2967..3680
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     misc_feature    3681..4217
                     /label=end of CLN2 ORF (PEST sequence)
                     /note="end of CLN2 ORF"
     misc_feature    4226..4237
                     /label=ADH1 end of ORF
                     /note="ADH1 end of ORF"
     misc_feature    4238..4427
                     /label=ADH1 3prime UTR
                     /note="ADH1 3prime UTR"
     terminator      4238..4425
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4493..4509)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4535..4553)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4563..4579)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4724..5179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(5310..6113)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6114..6329)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc
     2281 accgcggtgg cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tcctttggat
     2341 ggacgcaaag aagtttaata atcatattac atggcattac caccatatac atatccatat
     2401 ctaatcttac ttatatgttg tggaaatgta aagagcccca ttatcttagc ctaaaaaaac
     2461 cttctctttg gaactttcag taatacgctt aactgctcat tgctatattg aagtacggat
     2521 tagaagccgc cgagcgggcg acagccctcc gacggaagac tctcctccgt gcgtcctcgt
     2581 cttcaccggt cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata
     2641 aagattctac aatactagct tttatggtta tgaagaggaa aaattggcag taacctggcc
     2701 ccacaaacct tcaaattaac gaatcaaatt aacaaccata ggatgataat gcgattagtt
     2761 ttttagcctt atttctgggg taattaatca gcgaagcgat gatttttgat ctattaacag
     2821 atatataaat ggaaaagctg cataaccact ttaactaata ctttcaacat tttcagtttg
     2881 tattacttct tattcaaatg tcataaaagt atcaacaaaa aattgttaat atacctctat
     2941 actttaacgt caaggagtta attaacatgt ctaaaggtga agaattattc actggtgttg
     3001 tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct gtctccggtg
     3061 aaggtgaagg tgatgctact tacggtaaat tgaccttaaa attgatttgt actactggta
     3121 aattgccagt tccatggcca accttagtca ctactttagg ttatggtttg caatgttttg
     3181 ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg ccagaaggtt
     3241 atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc agagctgaag
     3301 tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt gattttaaag
     3361 aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac aatgtttaca
     3421 tcactgctga caaacaaaag aatggtatca aagctaactt caaaattaga cacaacattg
     3481 aagatggtgg tgttcaatta gctgaccatt atcaacaaaa tactccaatt ggtgatggtc
     3541 cagtcttgtt accagacaac cattacttat cctatcaatc tgccttatcc aaagatccaa
     3601 acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt attacccatg
     3661 gtattgatga attgtacaaa gcatccaact tgaacatttc gagaaagctt accatatcaa
     3721 ccccatcatg ctctttcgaa aattcaaata gcacatccat tccttcgccc gcttcctcat
     3781 ctcaaagcca cactccaatg agaaacatga gctcactctc tgataacagc gttttcagcc
     3841 ggaatatgga acaatcatca ccaatcactc caagtatgta ccaatttggt cagcagcagt
     3901 caaacagtat atgtggtagc accgttagtg tgaatagtct ggtgaataca aataacaaac
     3961 aaaggatcta cgaacaaatc acgggtccta acagcaataa cgcaaccaat gattatattg
     4021 atttgctaaa cctaaatgag tctaacaagg aaaaccaaaa tcccgcaacg gcgcattacc
     4081 tcaatggggg cccacccaag acaagcttca ttaaccatgg aatgttcccc tcgccaactg
     4141 ggaccataaa tagcggtaaa tctagcagtg cctcatcttt aatttctttt ggtatgggca
     4201 atacccaagt aatatagggc gcgccacttc taaataagcg aatttcttat gatttatgat
     4261 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     4321 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     4381 tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat ccgctaggga
     4441 taacagggta atatagatcc cccgggctgc aggaattcga tatcaagctt atcgataccg
     4501 tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     4561 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4621 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4681 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc
     4741 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4801 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4861 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4921 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4981 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5041 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5101 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5161 atattaacgt ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5221 cacaccgcat agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt
     5281 atacatgcat ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat
     5341 gcttcccagc ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca
     5401 aatagtcctc ttccaacaat aataatgtca gatcctgtag agaccacatc atccacggtt
     5461 ctatactgtt gacccaatgc gtctcccttg tcatctaaac ccacaccggg tgtcataatc
     5521 aaccaatcgt aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca
     5581 aaatctttgt cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg
     5641 gagcccttgc atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct
     5701 tctgccgcct gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta
     5761 atgtctgccc attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta
     5821 ccaatgtcag caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc
     5881 tttagcggct taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt
     5941 agtaaacaaa ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga
     6001 acatccaatg aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca
     6061 gcaacaggac taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat
     6121 cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat
     6181 gtttcttcaa cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt
     6241 tcttccttct gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa
     6301 aaagaataaa aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg cactctcagt
     6361 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6421 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6481 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcga
//
