LOCUS       62056_22945        8177 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 8177)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        335..742
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     rep_origin      complement(1272..1727)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1872..1888
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1898..1916
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1935..2182)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2185..2201
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2235..2251)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2257..2900)
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     promoter        2920..2938
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             3379..3642
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAE"
     terminator      4371..4535
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        complement(4575..4593)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4614..4630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4638..4654)
                     /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)."
     promoter        complement(4662..4692)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4707..4728)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5016..5604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5778..6635)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6636..6740)
                     /label=AmpR promoter
     rep_origin      complement(6767..8109)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgccacac accatagctt caaaatgttt
      361 ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa
      421 aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta
      481 cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa
      541 aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt tttgattttt
      601 ttctctttcg atgacctccc attgatattt aagttaataa acggtcttca atttctcaag
      661 tttcagtttc atttttcttg ttctattaca acttttttta cttcttgctc attagaaaga
      721 aagcatagca atctaatcta agggcgagct cgaattcgaa atgaccgacc aagcgacgcc
      781 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
      841 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
      901 cttcgcccac cccgggctcg atcccctcgc gagttggttc agctgctgcc tgaggctgga
      961 cgacctcgcg gagttctacc ggcagtgcaa atccgtcggc atccaggaaa ccagcagcgg
     1021 ctatccgcgc atccatgccc ccgaactgca ggagtgggga ggcacgatgg ccgctttggt
     1081 cgacccggac gggacgctcc tgcgcctgat acagaacgaa ttgcttgcag gcatctcatg
     1141 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     1201 attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1261 accgcatcag gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1321 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1381 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1441 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1501 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1561 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     1621 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     1681 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcgc gccattcgcc
     1741 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     1801 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     1861 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     1921 attgggtacc ggccgcaaat taaagccttc gagcgtccca aaaccttctc aagcaaggtt
     1981 ttcagtataa tgttacatgc gtacacgcgt ctgtacagaa aaaaaagaaa aatttgaaat
     2041 ataaataacg ttcttaatac taacataact ataaaaaaat aaatagggac ctagacttca
     2101 ggttgtctaa ctccttcctt ttcggttaga gcggatgtgg ggggagggcg tgaatgtaag
     2161 cgtgacataa ctaattacat gactcgaggt cgacggtatc gataagcttg atatcgaatt
     2221 cctgcagccc gggggatcca ctagttctag aatccgtcga aactaagttc tggtgtttta
     2281 aaactaaaaa aaagactaac tataaaagta gaatttaaga agtttaagaa atagatttac
     2341 agaattacaa tcaataccta ccgtctttat atacttatta gtcaagtagg ggaataattt
     2401 cagggaactg gtttcaacct tttttttcag ctttttccaa atcagagaga gcagaaggta
     2461 atagaaggtg taagaaaatg agatagatac atgcgtgggt caattgcctt gtgtcatcat
     2521 ttactccagg caggttgcat cactccattg aggttgtgcc cgttttttgc ctgtttgtgc
     2581 ccctgttctc tgtagttgcg ctaagagaat ggacctatga actgatggtt ggtgaagaaa
     2641 acaatatttt ggtgctggga ttcttttttt ttctggatgc cagcttaaaa agcgggctcc
     2701 attatattta gtggatgcca ggaataaact gttcacccag acacctacga tgttatatat
     2761 tctgtgtaac ccgcccccta ttttgggcat gtacgggtta cagcagaatt aaaaggctaa
     2821 ttttttgact aaataaagtt aggaaaatca ctactattaa ttatttacgt attctttgaa
     2881 atggcgagta ttgataatga taaactgagc tcggcgcgca attaaccctc actaaaggga
     2941 acaaaagctg gagctcatag cttcaaaatg tttctactcc ttttttactc ttccagattt
     3001 tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt
     3061 cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa
     3121 aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt
     3181 tctttttctt gaaaattttt tttttgattt ttttctcttt cgatgacctc ccattgatat
     3241 ttaagttaat aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta
     3301 caactttttt tacttcttgc tcattagaaa gaaagcatag caatctaatc taagttttct
     3361 agaactagcg cggccgctat gagatttcct tcaattttta ctgctgtttt attcgcagca
     3421 tcctccgcat tagctgctcc agtcaacact acaacagaag atgaaacggc acaaattccg
     3481 gctgaagctg tcatcggtta ctcagattta gaaggggatt tcgatgttgc tgttttgcca
     3541 ttttccaaca gcacaaataa cgggttattg tttataaata ctactattgc cagcattgct
     3601 gctaaagaag aaggggtatc tctcgagaag agagaggctg aaaacggtat cttcaacact
     3661 agattgtcta gaactatcgg ttacactgtt aagaagacta ctgttagaac tccatcttgg
     3721 aacgttgaca tgatgagatt caacatcaac gacttcttgc caccaggtgg tggttctaac
     3781 ccattgactg ttccattcga atactacaga atcagaaagg ttaaggttga attctggcca
     3841 tgttctccaa tcactcaagg tgacagaggt gttggttcta ctgctgttat cttggacgac
     3901 aacttcgtta ctaaggctaa cgctttgact tacgacccat acgttaacta ctcttctaga
     3961 cacactatca ctcaaccatt ctactaccac tctagatact tcactccaaa gccagttttg
     4021 gacagaacta tcgactactt ccaaccaaac aacaagagaa accaattgtg gttgagattg
     4081 caaactactg gtaacgttga ccacgttggt ttgggtactg ctttcgaaaa ctctatctac
     4141 gaccaagact acaacatcag aatcactatg tacgttcaat tcagagaatt caacttgaag
     4201 gacccaccat tgaagccata accgcggaga tctttaatta acaattcttc gccagaggtt
     4261 tggtcaagtc tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga
     4321 tggaaaaggg tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct
     4381 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt
     4441 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg
     4501 tcaggttgct ttctcagtat agcatgaggt cgctccaatt cagctggcgt aatagcgagc
     4561 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4621 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc
     4681 ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc
     4741 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4801 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4921 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5641 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5701 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5761 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5821 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5881 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5941 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6001 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6061 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6121 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6181 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6241 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6301 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6361 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6421 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6481 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6541 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6601 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6661 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6721 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaacg aagcatctgt
     6781 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     6841 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     6901 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     6961 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     7021 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     7081 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     7141 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     7201 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     7261 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     7321 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     7381 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     7441 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     7501 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     7561 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     7621 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     7681 ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     7741 actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc
     7801 ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc
     7861 tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta
     7921 aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga
     7981 tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc
     8041 tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga
     8101 tattggatca tactaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     8161 cacgaggccc tttcgtc
//
