LOCUS       62056_23085        6907 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6907)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        475..493
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             804..1520
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1602..1619
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1653..1900
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(2148..2736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2910..3767)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(3866..4666)
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4667..4887)
                     /label=URA3 promoter
     rep_origin      complement(5486..6366)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6377..6890)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
      481 gactcactat agggaatatt aagcttggta ccgccaccat gagatttcct tcaattttta
      541 ctgcagtttt attcgcagca tcctccgcat tagctgctcc agtcaacact acaacagaag
      601 atgaaacggc acaaattccg gctgaagctg tcatcggtta ctcagattta gaaggggatt
      661 tcgatgttgc tgttttgcca ttttccaaca gcacaaataa cgggttattg tttataaata
      721 ctactattgc cagcattgct gctaaagaag aaggggtatc tctcgagaaa agagaggctg
      781 aagcttacgt agaattccct aggatggtga gcaagggcga ggagctgttc accggggtgg
      841 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      901 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      961 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1021 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1081 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1141 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1201 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1261 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1321 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1381 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1441 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1501 gcatggacga gctgtacaag gcggccgcga attctagagg gcccttcgaa ggtaagccta
     1561 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac catcaccatt
     1621 gagtttaaac ccgctgatcc tagagggccg catcatgtaa ttagttatgt cacgcttaca
     1681 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     1741 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     1801 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     1861 gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctgcggc cctgcattaa
     1921 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1981 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2041 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2101 ggccagcaaa agcccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2161 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2221 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2281 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2341 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2401 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2461 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2521 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2581 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2641 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2701 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2761 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2821 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2881 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2941 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3001 atacgggagc gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3061 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3121 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3181 agttcgccag ttaatagttt gcgcaacgtt gttggcattg ctacaggcat cgtggtgtca
     3241 ctctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3301 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3361 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3421 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3481 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taatagtgta
     3541 tcacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3601 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3661 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3721 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3781 caatgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg
     3841 catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc
     3901 agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc
     3961 ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg gttctatact
     4021 gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat
     4081 cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt
     4141 tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct
     4201 tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg
     4261 cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg
     4321 cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt
     4381 cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat gcctttagcg
     4441 gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac
     4501 aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca
     4561 atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag
     4621 gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt
     4681 tcctgcaggt ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt
     4741 caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct
     4801 tctgttcgga gattaccgaa tcaaaaaaat ttcaaagaaa ccgaaatcaa aaaaaagaat
     4861 aaaaaaaaaa tgatgaattg aattgaaaag ctagcttatc gatgataagc tgtcaaagat
     4921 gagaattaat tccacggact atagactata ctagatactc cgtctactgt acgatacact
     4981 tccgctcagg tccttgtcct ttaacgaggc cttaccactc ttttgttact ctattgatcc
     5041 agctcagcaa aggcagtgtg atctaagatt ctatcttcgc gatgtagtaa aactagctag
     5101 accgagaaag agactagaaa tgcaaaaggc acttctacaa tggctgccat cattattatc
     5161 cgatgtgacg ctgcagcttc tcaatgatat tcgaatacgc tttgaggaga tacagcctaa
     5221 tatccgacaa actgttttac agatttacga tcgtacttgt tacccatcat tgaattttga
     5281 acatccgaac ctgggagttt tccctgaaac agatagtata tttgaacctg tataataata
     5341 tatagtctag cgctttacgg aagacaatgt atgtatttcg gttcctggag aaactattgc
     5401 atctattgca taggtaatct tgcacgtcgc atccccggtt cattttctgc gtttccatct
     5461 tgcacttcaa tagcatatct ttgttaacga agcatctgtg cttcattttg tagaacaaaa
     5521 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     5581 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     5641 caaaaatgca acgcgacgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     5701 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     5761 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     5821 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     5881 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     5941 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     6001 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     6061 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     6121 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     6181 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     6241 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     6301 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     6361 cgcaatggga agctccaccc cggttgataa tcagaaaagc cccaaaaaca ggaagattgt
     6421 ataagcaaat atttaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     6481 ttaaatcagc tcatttttta acgaatagcc cgaaatcggc aaaatccctt ataaatcaaa
     6541 agaatagacc gagatagggt tgagtgttgt tccagtttcc aacaagagtc cactattaaa
     6601 gaacgtggac tccaacgtca aagggcgaaa aagggtctat cagggcgatg gcccactacg
     6661 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcag taaatcggaa
     6721 gggtaaacgg atgcccccat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6781 ggaagggaag aaagcgaaag gagcgggggc tagggcggtg ggaagtgtag gggtcacgct
     6841 gggcgtaacc accacacccg ccgcgcttaa tggggcgcta cagggcgcgt ggggatgatc
     6901 cactagt
//
