LOCUS       40924_43583        8414 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTL80, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8414)
  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 8414)
  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 8414)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8414)
  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..8414
                     /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            3637..5212
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin
                     resistance"
     promoter        complement(6005..6402)
                     /label=LEU2 promoter
     promoter        6508..6612
                     /label=AmpR promoter
     CDS             6613..7470
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7644..8232
                     /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 atgtacagtt
     3601 tagcttgcct tgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     3661 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     3721 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     3781 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     3841 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     3901 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     3961 acatccgaac ataaacaacc atgggtaaaa agcctgaact caccgcgacg tctgtcgaga
     4021 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     4081 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     4141 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     4201 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
     4261 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     4321 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     4381 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     4441 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     4501 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     4561 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     4621 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     4681 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     4741 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     4801 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     4861 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     4921 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     4981 ccagcactcg tccgagggca aaggaataat cagtactgac aataaaaaga ttcttgtttt
     5041 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     5101 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     5161 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     5221 tcgatactaa cgccgccatc cgatatcacc aaacatgttg ctggtgatta taataccatt
     5281 taggtgggtt gggttcttaa ctaggatcat ggcggcagaa tcaatcaatt gatgttgaac
     5341 cttcaatgta ggaaattcgt tcttgatggt ttcctccaca gtttttctcc ataatcttga
     5401 agaggccaaa acattagctt tatccaagga ccaaataggc aatggtggct catgttgtag
     5461 ggccatgaaa gcggccattc ttgtgattct ttgcacttct ggaacggtgt attgttcact
     5521 atcccaagcg acaccatcac catcgtcttc ctttctctta ccaaagtaaa tacctcccac
     5581 taattctctg acaacaacga agtcagtacc tttagcaaat tgtggcttga ttggagataa
     5641 gtctaaaaga gagtcggatg caaagttaca tggtcttaag ttggcgtaca attgaagttc
     5701 tttacggatt tttagtaaac cttgttcagg tctaacacta cctgtacccc atttaggacc
     5761 acccacagca cctaacaaaa cggcatcaac cttcttggag gcttccagcg cctcatctgg
     5821 aagtgggaca cctgtagcat cgatagcagc accaccaatt aaatgatttt cgaaatcgaa
     5881 cttgacattg gaacgaacat cagaaatagc tttaagaacc ttaatggctt cggctgtgat
     5941 cttgaccaac gtggtcacct ggcaaaacga cgatcttctt aggggcagac ataggggcag
     6001 acattagaat gtatatcctt gaaatatata tatatattgc tgaaatgtaa aaggtaagaa
     6061 aagttagaat aagacgattg ctaaccacct attggaaaaa acaataggtc cttaaataat
     6121 attgtcaact tcaagtattg tgatgcaagc atttagtcat gaacgcttct ctattctata
     6181 tgaaaagccg gttccggcct ctcacctttc ctttttctcc caatttttca gttgaaaaag
     6241 gtatatgcgc ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgtctgtg
     6301 cgatagcgcc cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt gctaagagat
     6361 tcgaactctt gcatcttacg atacctgagt attcccacag ttaattcttg aagacgaaag
     6421 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6481 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7141 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7321 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     7801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8281 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     8341 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     8401 gaggaagcgg aaga
//
