LOCUS       62056_10840        5403 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 5403)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..67
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    75..91
                     /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)."
     sig_peptide     112..174
                     /label=OmpA signal peptide
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
     CDS             205..228
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     terminator      439..525
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      617..644
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        663..754
                     /label=AmpR promoter
     CDS             755..1612
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1786..2374
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2621..3072
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(3445..3618)
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     protein_bind    complement(3638..3654)
                     /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(3662..3692)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3707..3728)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3744..4823)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4824..4901)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
ORIGIN
        1 catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat catcggctcg
       61 tataatgtgt ggaattgtga gcggataaca atttcacaca ggagatatca tatgaaaaag
      121 acagctatcg cgattgcagt ggcactggct ggtttcgcta ccgttgcgca agcttctcga
      181 gaattcccgg gtaccagatc tgtcgactac aaggacgatg acgacaagtg aagatcgatc
      241 tctcgatcga gtgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
      301 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
      361 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
      421 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
      481 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
      541 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
      601 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
      661 cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      721 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      781 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
      841 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
      901 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
      961 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     1021 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1081 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1141 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1201 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1261 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1321 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1381 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1441 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1501 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1561 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1621 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1681 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1741 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1801 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1861 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1921 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1981 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2041 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2101 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2161 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2221 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2281 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2341 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2401 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2461 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2521 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2581 tacgcatctg tgcggtattt cacaccgcat acagatcctg acgcgccctg tagcggcgca
     2641 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2701 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2761 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2821 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2881 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2941 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3001 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3061 ttaacgttta caggatctgt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3121 ttaagccagt atacactccg ctatcgctac gtgactgcct cgacctgcag caattccaac
     3181 gccatcaaaa ataattcgcg tctggccttc ctgtagccag ctttcatcaa cattaaatgt
     3241 gagcgagtaa caacccgtcg gattctccgt gggaacaaac ggcggattga ccgtaatggg
     3301 ataggtcacg ttggtgtaga tgggcgcatc gtaaccgtgc atctgccagt ttgaggggac
     3361 gacgacagta tcggcctcag gaagatcgca ctccagccag ctttccggca ccgcttctgg
     3421 tgccggaaac caggcaaagc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3481 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3541 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3601 atccgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3661 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3721 ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3781 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg
     3841 gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga
     3901 gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg
     3961 gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag
     4021 atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc
     4081 tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt
     4141 tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga
     4201 ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat
     4261 gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt
     4321 cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca
     4381 agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc
     4441 agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct
     4501 ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga
     4561 tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag
     4621 gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga
     4681 atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg
     4741 ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca
     4801 tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat
     4861 catgccatac cgcgaaaggt tttgcaccat tccatggtgt cggaattgct gcaggtcgag
     4921 ggggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4981 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     5041 aggttttcac cgtcatcacc gaaacgcgcg aggcaggatg gcgcccaaca gtcccccggc
     5101 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     5161 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     5221 ggtgatgccg gccacgatgc gtccggcgta gaggatccgg gcttatcgac tgcacggtgc
     5281 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     5341 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     5401 cga
//
