LOCUS       62056_13490       10115 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 10115)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      355..973
                     /label=LAC4 terminator
                     /note="transcription terminator for Kluyveromyces lactis
                     LAC4"
     promoter        974..1676
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1697..3340
                     /codon_start=1
                     /label=amdS
                     /note="acetamidase from Aspergillus nidulans"
                     /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGIL
                     SEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALAQA
                     RELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTMLRK
                     AGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGVIGVG
                     TDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVEDLRLFT
                     KSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPPILRGVE
                     TTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAIPNIKDLL
                     NPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPTAAVRHDQF
                     RYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYDPEAYHGAPV
                     AVQVIGRRLSEERTLAIAEEVGKLLGNVVTP"
     promoter        3979..4635
                     /label=LAC4 promoter (5')
                     /note="5' portion of the Kluyveromyces lactis LAC4
                     promoter"
     CDS             4638..5351
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="MGKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      5414..5661
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(5676..5694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5756..5786)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5801..5822)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6110..6698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6872..7729)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7730..7834)
                     /label=AmpR promoter
     rep_origin      7861..8316
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8487..10000
                     /label=LAC4 promoter (3')
                     /note="3' portion of a modified Kluyveromyces lactis LAC4 
                     promoter"
     protein_bind    10021..10037
                     /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)."
ORIGIN
        1 aagcttgaaa aaaatgaaat tctctactat attagccgca tctactgctt taatttccgt
       61 tgttatggct gctccagttt ctaccgaaac tgacatcgac gatcttccaa tatcggttcc
      121 agaagaagcc ttgattggat tcattgactt aaccggggat gaagtttcct tgttgcctgt
      181 taataacgga acccacactg gtattctatt cttaaacacc accatcgctg aagctgcttt
      241 cgctgacaag gatgatctcg agaaaagaga ggctgaagct agaagagcta gatctcctag
      301 gggtaccgtc gacggcgcgc ctgcggccgc ttaattaagg ccttgaatcg agaatttata
      361 cttagataag tatgtactta caggtatatt tctatgagat actgatgtat acatgcatga
      421 taatatttaa acggttatta gtgccgattg tcttgtgcga taatgacgtt cctatcaaag
      481 caatacactt accacctatt acatgggcca agaaaatatt ttcgaacttg tttagaatat
      541 tagcacagag tatatgatga tatccgttag attatgcatg attcattcct acaacttttt
      601 cgtagcataa ggattaatta cttggatgcc aataaaaaaa aaaaacatcg agaaaatttc
      661 agcatgctca gaaacaattg cagtgtatca aagtaaaaaa aagattttca ctacatgttc
      721 cttttgaaga aagaaaatca tggaacatta gatttacaaa aatttaacca ccgctgatta
      781 acgattagac cgttaagcgc acaacaggtt attagtacag agaaagcatt ctgtggtgtt
      841 gccccggact ttcttttgcg acataggtaa atcgaatacc atcatactat cttttccaat
      901 gactccctaa agaaagactc ttcttcgatg ttgtatacgt tggagcatag ggcaagaatt
      961 gtggcttgag atcatccttt tgttgtttcc gggtgtacaa tatggacttc ctcttttctg
     1021 gcaaccaaac ccatacatcg ggattcctat aataccttcg ttggtctccc taacatgtag
     1081 gtggcggagg ggagatatac aatagaacag ataccagaca agacataatg ggctaaacaa
     1141 gactacacca attacactgc ctcattgatg gtggtacata acgaactaat actgtagccc
     1201 tagacttgat agccatcatc atatcgaagt ttcactaccc tttttccatt tgccatctat
     1261 tgaagtaata ataggcgcat gcaacttctt ttcttttttt ttcttttctc tctcccccgt
     1321 tgttgtctca ccatatccgc aatgacaaaa aaatgatgga agacactaaa ggaaaaaatt
     1381 aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa
     1441 gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga gcaacggtat
     1501 acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt cttgctatca
     1561 agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc tcgttccctt
     1621 tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac aatcaaggaa
     1681 ttccggatcc gccaccatgc ctcaatcctg ggaagaactg gccgctgata agcgcgcccg
     1741 cctcgcaaaa accatccctg atgaatggaa agtccagacg ctgcctgcgg aagacagcgt
     1801 tattgatttc ccaaagaaat cggggatcct ttcagaggcc gaactgaaga tcacagaggc
     1861 ttccgctgcg gatcttgtgt ccaagctggc ggccggagag ttgacctcgg tggaagttac
     1921 gctagcattc tgtaaacggg cagcaatcgc ccagcagtta acaaactgcg cccacgagtt
     1981 cttccctgac gccgctctcg cgcaggcaag ggaactcgat gaatactacg caaagcacaa
     2041 gagacccgtt ggtccactcc atggcctccc catctctctc aaagaccagc ttcgagtcaa
     2101 gggctacgaa acatcaatgg gctacatctc atggctaaac aagtacgacg aaggggactc
     2161 ggttctgaca accatgctcc gcaaagccgg tgccgtcttc tacgtcaaga cctctgtccc
     2221 gcagaccctg atggtctgcg agacagtcaa caacatcatc gggcgcaccg tcaacccacg
     2281 caacaagaac tggtcgtgcg gcggcagttc tggtggtgag ggtgcgatcg ttgggattcg
     2341 tggtggcgtc atcggtgtag gaacggatat cggtggctcg attcgagtgc cggccgcgtt
     2401 caacttcctg tacggtctaa ggccgagtca tgggcggctg ccgtatgcaa agatggcgaa
     2461 cagcatggag ggtcaggaga cggtgcacag cgttgtcggg ccgattacgc actctgttga
     2521 ggacctccgc ctcttcacca aatccgtcct cggtcaggag ccatggaaat acgactccaa
     2581 ggtcatcccc atgccctggc gccagtccga gtcggacatt attgcctcca agatcaagaa
     2641 cggcgggctc aatatcggct actacaactt cgacggcaat gtccttccac accctcctat
     2701 cctgcgcggc gtggaaacta ccgtcgccgc actcgccaaa gccggtcaca ccgtgacccc
     2761 gtggacgcca tacaagcacg atttcggcca cgatctcatc tcccatatct acgcggctga
     2821 cggcagcgcc gacgtaatgc gcgatatcag tgcatccggc gagccggcga ttccaaatat
     2881 caaagaccta ctgaacccga acatcaaagc tgttaacatg aacgagctct gggacacgca
     2941 tctccagaag tggaattacc agatggagta ccttgagaaa tggcgggagg ctgaagaaaa
     3001 ggccgggaag gaactggacg ccatcatcgc gccgattacg cctaccgctg cggtacggca
     3061 tgaccagttc cggtactatg ggtatgcctc tgtgatcaac ctgctggatt tcacgagcgt
     3121 ggttgttccg gttacctttg cggataagaa catcgataag aagaatgaga gtttcaaggc
     3181 ggttagtgag cttgatgccc tcgtgcagga agagtatgat ccggaggcgt accatggggc
     3241 accggttgca gtgcaggtta tcggacggag actcagtgaa gagaggacgt tggcgattgc
     3301 agaggaagtg gggaagttgc tgggaaatgt ggtgactcca tagcccgggg ggggctcgat
     3361 cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg
     3421 cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc
     3481 gaactgcagg agtggggagg cacgatggcc gctttggtcg atctagatta cgtggaagaa
     3541 aggtagtaaa agtagtagta taagtagtaa aaagaggtaa aaagagaaaa ccggctacat
     3601 actagagaag cacgtacaca aaaactcata ggcacttcat catacgacag tttcttgatg
     3661 cattataata gtgtattaga tattttcaga aatatgcata gaacctcctc ttgcctttac
     3721 tttttataca tagaacattg gcagatttac ttacactact ttgtttctac gccatttctt
     3781 ttgttttcaa cacttagaca agttgttgag aaccggacta ctaaaaagca atgttcccac
     3841 tgaaaatcat gtacctgcag gataataacc ccctaattct gcatcgatcc agtatgtttt
     3901 tttttctcta ctcattttta cctgaagata gagcttctaa aacaaaaaaa atcagtgatt
     3961 acatgcatat tgtgtgttct agtaaccaaa ggaaaggaac agatagataa aattccgaga
     4021 ctgtcaaatt aggttttttt cttttttttt ggcgggagtc agtgggccga aatatgttct
     4081 tggcctagaa cttaatctgg tttgatcatg ccaatacttg cctgagtgcc cgactttttg
     4141 cccaccctct tgccttctgt catccttcaa aacccacctg ttttccagcc gtatcttcgc
     4201 tcgcatctac acatactgtg ccatatcttg tgtgtagccg gacgtgacta tgaccaaaaa
     4261 caaacaagga gaactgttcg ccgatttgta acactcctgc atccatccaa gtgggtatgc
     4321 gctatgcaat gttaagctag gtcaggtcag accaggtcca aggacagcaa cttgactgta
     4381 tgcaaccttt accatctttg cacagaacat acttgtagct agctagttac acttatggac
     4441 cgaaaaggca ccccaccatg tctgtccggc tttagagtac ggccgcagac cgctgatttg
     4501 ccttgccaag cagtagtcac aatgcatcgc atgagcacac gggcacgggc acgggcacag
     4561 gaaccattgg caaaaatacc agatacacta taccgacgta tatcaagccc aagtttaaaa
     4621 ttcctaaatt tccgcggatg ggtaagggag aagaactttt cactggagtt gtcccaattc
     4681 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     4741 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aagcttcctg
     4801 ttccttggcc aacacttgtc actactctta cttatggtgt tcaatgcttt tcaagatacc
     4861 cagatcatat gaagcggcac gacttcttca agagcgccat gcctgaggga tacgtgcagg
     4921 agaggaccat cttcttcaag gacgacggga actacaagac acgtgctgaa gtcaagtttg
     4981 agggagacac cctcgtcaac agaatcgagc ttaagggaat cgatttcaag gaggacggaa
     5041 acatcctcgg ccacaagttg gaatacaact acaactccca caacgtatac atcatggcag
     5101 acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa
     5161 gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     5221 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga
     5281 gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg
     5341 aactatacaa ataagaattc tgcagatatc catcacactg gcggccgctc gagcatgcat
     5401 ctagagggcc gcatcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     5461 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     5521 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     5581 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     5641 cgctcgaagg ctttaatttg cctacttttc aattccctat agtgagtcgt attaaattcg
     5701 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5821 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5881 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6061 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6121 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6181 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6241 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6301 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6361 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6421 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6481 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6541 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6601 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6661 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6721 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7861 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7921 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7981 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     8041 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     8101 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     8161 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     8221 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     8281 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa
     8341 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     8401 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     8461 aacgacggcc agtgccaagc tcccgcgggg atcgactcat aaaatagtaa ccttctaatg
     8521 cgtatctatt gactaccaac cattagtgtg gttgcagaag gcggaattct cccttcttcg
     8581 aattcagctt gctttttcat tttttatttt ccatttttca gtttttgttt gtgtcgaatt
     8641 tagccagttg cttctccaag atgaaaaaaa cccctgcgca gtttctgtgc tgcaagatcc
     8701 taatcgactt ttccaccccc cacaaaagta aatgttcttt tgttacattc gcgtgggtag
     8761 ctagctcccc gaatcttcaa aggacttagg gactgcacta catcagagtg tgttcacctg
     8821 gtttgctgcc tggtttgaaa gaaaagagca gggaactcgc gggttcccgg cgaataatca
     8881 tgcgatagtc ctttggcctt ccaagtcgca tgtagagtag acaacagaca gggagggcag
     8941 gaaggatctt tcactgagat cctgtatctt gttgggtaag tcggatgaaa ggggaatcgt
     9001 atgagattgg agaggatgcg gaagaggtaa cgccttttgt taacttgttt aattattatg
     9061 gggcaggcga gagggggagg aatgtatgtg tgtgaggcgg gcgagacgga gccatccagg
     9121 ccaggtagaa atagagaaag ccgaatgtta gacaatatgg cagcgtagta gagtaggtag
     9181 gtaggcaagt actgctagca aagaggagaa gggtaagctc actcttcgca ttccacaccg
     9241 ttagtgtgtc agtttgaaca aaaaaacaat catcatacca attgatggac tgtggactgg
     9301 cttttggaac ggcttttcgg actgcgatta ttcgtgagga atcaaggtag gaatttggtc
     9361 atatttacgg acaacagtgg gtgattccca tatggagtag gaaaacgaga tcatggtatc
     9421 ctcagatatg ttgcggaatt ctgttcaccg caaagttcag ggtgctctgg tgggtttcgg
     9481 ttggtctttg ctttgcttct cccttgtctt gcatgttaat aatagcctag cctgtgagcc
     9541 gaaacttagg gtaggcttag tgttggaacg tacatatgta tcacgttgac ttggtttaac
     9601 caggcgacct ggtagccagc catacccaca cacgtttttt gtatcttcag tatagttgtg
     9661 aaaagtgtag cggaaatttg tggtccgagc aacagcgtct ttttctagta gtgcggtcgg
     9721 ttacttggtt gacattggta tttggacttt gttgctacac cattcactac ttgaagtcga
     9781 gtgtgaaggg tatgatttct agtggtgaac acctttagtt acgtaatgtt ttcattgctg
     9841 ttttacttga gatttcgatt gagaaaaagg tatttaatag ctcgaatcaa tgtgagaaca
     9901 gagagaagat gttcttccct aactcgaaag gtatatgagg cttgtgtttc ttaggagaat
     9961 tattattctt ttgttatgtt gcgcttgtag ttggaaaagg tgaagagaca aaagctggaa
    10021 ttgtgagcgg ataacaagct caacacttga aatttaggaa agagcagaat ttggcaaaaa
    10081 aaataaaaaa aaaataaaca cacatactca tcgag
//
