LOCUS       40924_43588        8197 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTL85, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8197)
  AUTHORS   Baruffini E, Serafini F, Lodi T.
  TITLE     Construction and characterization of centromeric, episomal and 
            GFP-containing vectors for Saccharomyces cerevisiae prototrophic 
            strains
  JOURNAL   J. Biotechnol. 143 (4), 247-254 (2009)
  PUBMED    19683551
REFERENCE   2  (bases 1 to 8197)
  AUTHORS   Baruffini E, Serafini F, Lodi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2009) Department of Genetics, Biology of 
            Microorganisms, Anthropology, Evolution, University of Parma, Viale 
            Usberti 11/A, Parma 43100, Italy
REFERENCE   3  (bases 1 to 8197)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8197)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol."; date: "2009"; volume: "143"; issue: "4"; pages: 
            "247-254"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (18-MAR-2009) Department of Genetics, Biology of Microorganisms, 
            Anthropology, Evolution, University of Parma, Viale Usberti 11/A, 
            Parma 43100, Italy"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /label=lac operator
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(249..496)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(509..1222)
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     1225..1241
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1275..1291)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1292..1641)
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     complement(1686..1702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2139..3485)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     gene            3639..4995
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     promoter        complement(5788..6185)
                     /label=LEU2 promoter
     promoter        6291..6395
                     /label=AmpR promoter
     CDS             6396..7253
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7427..8015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcggccgc aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt
      301 ataatgttac atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat
      361 aacgttctta atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt
      421 ctaactcctt ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac
      481 ataactaatt acatgactcg accagttatt tgtacaattc atccatacca tgggtaatac
      541 cagcagcagt aacaaattct aacaagacca tgtggtctct cttttcgttt ggatctttgg
      601 ataaggcaga ttgagtggat aagtaatggt tgtctggtaa caagactgga ccatcaccaa
      661 ttggagtatt ttgttgataa tggtcagcta attgaacaga accatcttca atgttgtgtc
      721 taattttgaa gttaactttg ataccattct tttgtttgtc agccatgatg taaacattgt
      781 gagagttata gttgtattcc aatttgtgac ctaaaatgtt accatcttct ttaaaatcaa
      841 taccttttaa ttcgattcta ttaactaagg tatcaccttc aaacttgact tcagctctgg
      901 tcttgtagtt accgtcatct ttgaaaaaaa tagttctttc ttgaacataa ccttctggca
      961 tggcagactt gaaaaagtca tgttgtttca tatgatctgg gtatctagca aaacattgaa
     1021 caccataacc gaaagtagtg actaaggttg gccatggaac tggcaattta ccagtagtac
     1081 aaataaattt taaggtcaat ttaccgtaag tagcatcacc ttcaccttca ccggagacag
     1141 aaaatttgtg accattaaca tcaccatcta attcaaccaa aattgggaca acaccagtga
     1201 ataattcttc acctttagac atgtcgaggt cgacggtatc gataagcttg atatcgaatt
     1261 cctgcagccc gggggatcca ctagttctag agtatggatg ggggtaatag aattgtatct
     1321 atgtatctga cgaccctgta ttacacgaag gaaatagtag atgaaaagac aagagagcaa
     1381 gaaaaaggaa aaacttcctt tctaacagac gctttactta accttatata tatcttattt
     1441 ttttcatcga gcgtgttcaa ttggacaagg tgccaccttt tcgacacttc ggttattatg
     1501 ttacacagtt ttcatgagga tggcgccttg actaacttaa ttagtcattt gccaaccacc
     1561 acagttcccc aatatcgaca gcttcacgtg ccatttgcca ttttgcgatc atgtgacctc
     1621 aagagaaaaa gcaaaaaata atgagagcta agggattcga acccttgcat ccggagctcg
     1681 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     1741 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     1801 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     1861 ctccttacgc atctgtgcgg tatttcacac cgcatatatc ggatcgtact tgttacccat
     1921 cattgaattt tgaacatccg aacctgggag ttttccctga aacagatagt atatttgaac
     1981 ctgtataata atatatagtc tagcgcttta cggaagacaa tgtatgtatt tcggttcctg
     2041 gagaaactat tgcatctatt gcataggtaa tcttgcacgt cgcatccccg gttcattttc
     2101 tgcgtttcca tcttgcactt caatagcata tctttgttaa cgaagcatct gtgcttcatt
     2161 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     2221 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     2281 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     2341 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     2401 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     2461 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     2521 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     2581 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     2641 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     2701 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     2761 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     2821 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     2881 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     2941 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     3001 aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
     3061 aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
     3121 gctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc
     3181 gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg
     3241 tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg
     3301 cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt
     3361 atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata
     3421 tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt
     3481 ggatcgatcc gatgataagc tgtcaaacat gagaattaat tctaccctat gaacatattc
     3541 cattttgtaa tttcgtgtcg tttctattat gaatttcatt tataaagttt atgtacactg
     3601 tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc
     3661 tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt
     3721 agcccataca tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc
     3781 gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag
     3841 acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc
     3901 cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca
     3961 tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga ggccgcgatt
     4021 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     4081 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     4141 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     4201 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     4261 gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag aatatcctga
     4321 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     4381 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     4441 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     4501 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     4561 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     4621 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     4681 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     4741 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagtact
     4801 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
     4861 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc
     4921 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg
     4981 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccgatatc accaaacatg
     5041 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     5101 gaatcaatca attgatgttg aaccttcaat gtaggaaatt cgttcttgat ggtttcctcc
     5161 acagtttttc tccataatct tgaagaggcc aaaacattag ctttatccaa ggaccaaata
     5221 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     5281 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     5341 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     5401 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     5461 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     5521 ctacctgtac cccatttagg accacccaca gcacctaaca aaacggcatc aaccttcttg
     5581 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     5641 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     5701 accttaatgg cttcggctgt gatcttgacc aacgtggtca cctggcaaaa cgacgatctt
     5761 cttaggggca gacatagggg cagacattag aatgtatatc cttgaaatat atatatatat
     5821 tgctgaaatg taaaaggtaa gaaaagttag aataagacga ttgctaacca cctattggaa
     5881 aaaacaatag gtccttaaat aatattgtca acttcaagta ttgtgatgca agcatttagt
     5941 catgaacgct tctctattct atatgaaaag ccggttccgg cctctcacct ttcctttttc
     6001 tcccaatttt tcagttgaaa aaggtatatg cgcggcgacc tctgaaatta acaaaaaatt
     6061 tccagtcatc gaatttgtct gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa
     6121 aaaaataatg gttgctaaga gattcgaact cttgcatctt acgatacctg agtattccca
     6181 cagttaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6241 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6301 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6361 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6421 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6481 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6541 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6601 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6661 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6721 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6781 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6841 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6901 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6961 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7021 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7081 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7141 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7201 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7261 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7321 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7381 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7441 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7501 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7561 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7621 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7681 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7741 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7801 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7861 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7921 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7981 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8041 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8101 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8161 ccgagcgcag cgagtcagtg agcgaggaag cggaaga
//
