LOCUS       Exported                6155 bp ds-DNA     circular SYN 29-JUN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6155)
  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..6155
                     /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    2333..2583
                     /label=tetOpr from pCH001
     protein_bind    2377..2395
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2593..3843
                     /codon_start=1
                     /label=yEVenus-CLN2PEST
                     /note="yEVenus-CLN2PEST"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYKASNLNISRKLTISTPSCSFENSNSTSIPSPASSSQSHTPMRNM
                     SSLSDNSVFSRNMEQSSPITPSMYQFGQQQSNSICGSTVSVNSLVNTNNKQRIYEQITG
                     PNSNNATNDYIDLLNLNESNKENQNPATAHYLNGGPPKTSFINHGMFPSPTGTINSGKS
                     SSASSLISFGMGNTQVI"
     misc_feature    2593..3306
                     /label=yEVenus from pKT0090 (addgene), but no stop codon
                     /note="yEVenus as pulled from pKT0090 plasmid addgene, just
                     missing stop codon"
     CDS             2593..3306
                     /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    3307..3843
                     /label=end of CLN2 ORF (PEST sequence)
                     /note="end of CLN2 ORF"
     misc_feature    3852..3863
                     /label=ADH1 end of ORF
                     /note="ADH1 end of ORF"
     misc_feature    3864..4053
                     /label=ADH1 3prime UTR
                     /note="ADH1 3prime UTR"
     terminator      3864..4051
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4119..4135)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4161..4179)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4189..4205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4350..4805
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(4936..5739)
                     /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(5740..5955)
                     /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 tccagatccg
     2341 ctagggataa cagggtaata tagatcaatt cctcgatccc tatcagtgat agagagtcga
     2401 caaagtcgag tttctcgatc gagaccactg catgcatgtg ctctgtatgt atataaaact
     2461 cttgttttct tcttttctct aaatattctt tccttataca ttaggtcctt tgtagcataa
     2521 attactatac ttctatagac acgcaaacac aaatacacac actaaattac cggatcaatt
     2581 cggttaatta acatgtctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt
     2641 gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat
     2701 gctacttacg gtaaattgac cttaaaattg atttgtacta ctggtaaatt gccagttcca
     2761 tggccaacct tagtcactac tttaggttat ggtttgcaat gttttgctag atacccagat
     2821 catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga
     2881 actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt
     2941 gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt
     3001 ttaggtcaca aattggaata caactataac tctcacaatg tttacatcac tgctgacaaa
     3061 caaaagaatg gtatcaaagc taacttcaaa attagacaca acattgaaga tggtggtgtt
     3121 caattagctg accattatca acaaaatact ccaattggtg atggtccagt cttgttacca
     3181 gacaaccatt acttatccta tcaatctgcc ttatccaaag atccaaacga aaagagagac
     3241 cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat tgatgaattg
     3301 tacaaagcat ccaacttgaa catttcgaga aagcttacca tatcaacccc atcatgctct
     3361 ttcgaaaatt caaatagcac atccattcct tcgcccgctt cctcatctca aagccacact
     3421 ccaatgagaa acatgagctc actctctgat aacagcgttt tcagccggaa tatggaacaa
     3481 tcatcaccaa tcactccaag tatgtaccaa tttggtcagc agcagtcaaa cagtatatgt
     3541 ggtagcaccg ttagtgtgaa tagtctggtg aatacaaata acaaacaaag gatctacgaa
     3601 caaatcacgg gtcctaacag caataacgca accaatgatt atattgattt gctaaaccta
     3661 aatgagtcta acaaggaaaa ccaaaatccc gcaacggcgc attacctcaa tgggggccca
     3721 cccaagacaa gcttcattaa ccatggaatg ttcccctcgc caactgggac cataaatagc
     3781 ggtaaatcta gcagtgcctc atctttaatt tcttttggta tgggcaatac ccaagtaata
     3841 tagggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
     3901 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     3961 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
     4021 aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat
     4081 agatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga cctcgagggg
     4141 gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     4201 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     4261 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4321 gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc
     4381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4501 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4561 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4621 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4681 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4741 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4801 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg
     4861 taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac
     4921 ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc
     4981 ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc
     5041 aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc
     5101 caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc
     5161 ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct
     5221 cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga
     5281 caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt
     5341 caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc
     5401 tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa
     5461 ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac
     5521 tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt
     5581 gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc
     5641 acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg
     5701 atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca
     5761 ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     5821 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     5881 ggagattacc gaatcaaaaa aatttcaaag aaaccgaaat caaaaaaaag aataaaaaaa
     5941 aaatgatgaa ttgaattgaa aagctgtggt atggtgcact ctcagtacaa tctgctctga
     6001 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     6061 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6121 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
