LOCUS       62056_22920        8858 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 8858)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      69..1411
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1438..1542
                     /label=AmpR promoter
     CDS             1543..2400
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2574..3162
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3450..3471
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3486..3516
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3524..3540
                     /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     3548..3564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3585..3603
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(3643..3807)
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(4458..4721)
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAE"
     promoter        complement(5162..5180)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5200..5843
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             5861..6124
                     /codon_start=1
                     /label=alpha-factor secretion signal
                     /note="N-terminal secretion signal from S. cerevisiae 
                     alpha-factor"
                     /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
                     GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAE"
     primer_bind     complement(6658..6674)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6677..6924
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(6943..6961)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6971..6987)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7132..7587
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        complement(8117..8524)
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
     1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
     1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
     3481 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3601 agggaacaaa agctggagct cgctattacg ccagctgaat tggagcgacc tcatgctata
     3661 ctgagaaagc aacctgacct acaggaaaga gttactcaag aataagaatt ttcgttttaa
     3721 aacctaagag tcactttaaa atttgtatac acttattttt tttataactt atttaataat
     3781 aaaaatcata aatcataaga aattcgctta tttagaagtg tcaacaacgt atctaccaac
     3841 gatttgaccc ttttccatct tttcgtaaat ttctggcaag gtagacaagc cgacaacctt
     3901 gattggagac ttgaccaaac ctctggcgaa gaattgttaa ttaaagatct ccgcggttac
     3961 tctttctttc ttaatctgtc ttgaagattt ctagaggaag agaaactacg cataacttct
     4021 gctcttacta tttcccatgc acatggagaa tatgattttt cctgtagata taaggttaat
     4081 ctatgaaaat atttacgcac agccaatata gaatcttctt ctagtaatgg agtaccttct
     4141 aaacccgctt cttgcataac gcaagcttct agatctctta gctgttggtc caatccagta
     4201 cagaattgat gcaataatga ttcgttccat gcggcggctg aaccttctgt ggaaaacaat
     4261 tgaaatgtct gctgtaacat ttcgtgaact aaggccatag cctgagcttt ctgcacttga
     4321 ttaccgccaa aagcttcatg aggagatccg aaatctcttc tatgatccaa gcaagagaag
     4381 ggtgagattc tacgcatttg agcaagcaga cgcaaggctc ttgtatgagc tagtgaatgt
     4441 gtttgtggca agtcacattc agcctctctc ttctcgagag ataccccttc ttctttagca
     4501 gcaatgctgg caatagtagt atttataaac aataacccgt tatttgtgct gttggaaaat
     4561 ggcaaaacag caacatcgaa atccccttct aaatctgagt aaccgatgac agcttcagcc
     4621 ggaatttgtg ccgtttcatc ttctgttgta gtgttgactg gagcagctaa tgcggaggat
     4681 gctgcgaata aaacagcagt aaaaattgaa ggaaatctca tagcggccgc gctagttcta
     4741 gaaaacttag attagattgc tatgctttct ttctaatgag caagaagtaa aaaaagttgt
     4801 aatagaacaa gaaaaatgaa actgaaactt gagaaattga agaccgttta ttaacttaaa
     4861 tatcaatggg aggtcatcga aagagaaaaa aatcaaaaaa aaaattttca agaaaaagaa
     4921 acgtgataaa aatttttatt gcctttttcg acgaagaaaa agaaacgagg cggtctcttt
     4981 tttcttttcc aaacctttag tacgggtaat taacgacacc ctagaggaag aaagagggga
     5041 aatttagtat gctgtgcttg ggtgttttga agtggtacgg cgatgcgcgg agtccgagaa
     5101 aatctggaag agtaaaaaag gagtagaaac attttgaagc tatgagctcc agcttttgtt
     5161 ccctttagtg agggttaatt gcgcgccgag ctcagtttat cattatcaat actcgccatt
     5221 tcaaagaata cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat
     5281 tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa
     5341 tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg
     5401 gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt
     5461 ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg
     5521 cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa
     5581 tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac accttctatt
     5641 accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga
     5701 aattattccc ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg
     5761 taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt
     5821 aaaacaccag aacttagttt cgacggattc tagaactagt atgagatttc cttcaatttt
     5881 tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga
     5941 agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga
     6001 tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa
     6061 tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga agagagaggc
     6121 tgaagttcct gtacctgaag cattaagagc attgcccggt gctagaggtt gtcatctagc
     6181 ccaattcaag tctttgagtc cacaagcttt gcaagctttc aagagagcta aagatgcatt
     6241 tgaggaatca ttacttgaag actggaactg ttcatcaaga atttttccaa gaagtagaga
     6301 tttgaaacaa ctacaagttt gggaacgtcc agtagcattg gaagcggagg tagctctaac
     6361 cctgtccgta ttgggatctc tggcaaatag ttctttgcat agtagccttg accagccctt
     6421 gcatacttta agacatatac acgctcaatt acaagcatgc gttcccgcac aacctatggc
     6481 cggaccaagg ccaagaggta gacttcacca ttggcttcat agattacaag aagcgcaaaa
     6541 aaaggaacca cagtcatgtc tggaagctag tgtgatgttc aacttgtttc gtcttttgac
     6601 tagagatttg aagtgtgttg cttctggcga tctatgtgtt taagatatca agcttatcga
     6661 taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
     6721 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     6781 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     6841 ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg
     6901 ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt gagtcgtatt
     6961 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     7021 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     7081 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc gacgcgccct
     7141 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     7201 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     7261 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     7321 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     7381 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     7441 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     7501 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     7561 ttaacaaaat attaacgttt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     7621 cggtatttca caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg
     7681 agtttagtat acatgcattt acttataata cagtttttca tgagatgcct gcaagcaatt
     7741 cgttctgtat caggcgcagg agcgtcccgt ccgggtcgac caaagcggcc atcgtgcctc
     7801 cccactcctg cagttcgggg gcatggatgc gcggatagcc gctgctggtt tcctggatgc
     7861 cgacggattt gcactgccgg tagaactccg cgaggtcgtc cagcctcagg cagcagctga
     7921 accaactcgc gaggggatcg agcccggggt gggcgaagaa ctccagcatg agatccccgc
     7981 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     8041 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     8101 cgaattcgag ctcgccctta gattagattg ctatgctttc tttctaatga gcaagaagta
     8161 aaaaaagttg taatagaaca agaaaaatga aactgaaact tgagaaattg aagaccgttt
     8221 attaacttaa atatcaatgg gaggtcatcg aaagagaaaa aaatcaaaaa aaaaaatttt
     8281 caagaaaaag aaacgtgata aaaattttta ttgccttttt cgacgaagaa aaagaaacga
     8341 ggcggtctct tttttctttt ccaaaccttt agtacgggta attaacgaca ccctagagga
     8401 agaaagaggg gaaatttagt atgctgtgct tgggtgtttt gaagtggtac ggcgatgcgc
     8461 ggagtccgag aaaatctgga agagtaaaaa aggagtagaa acattttgaa gctatggtgt
     8521 gtggcatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg
     8581 ttcggagatt accgaatcaa aaaaatttca aagaaaccga aatcaaaaaa aagaataaaa
     8641 aaaaaatgat gaattgaatt gaaaagctgt ggtatggtgc actctcagta caatctgctc
     8701 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     8761 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     8821 gtgtcagagg ttttcaccgt catcaccgaa acgcgcga
//
