LOCUS       62056_8190        5023 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 5023)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     21..86
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     rep_origin      258..713
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        740..844
                     /label=AmpR promoter
     CDS             845..1702
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1876..2464
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2650..2792)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3201..3374)
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     protein_bind    complement(3394..3410)
                     /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(3418..3448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3463..3484)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3500..4579)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4580..4657)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     protein_bind    4709..4730
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4745..4775
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    4783..4799
                     /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        4854..4882
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
ORIGIN
        1 atcgatgctt aggaggtcat atgaaatacc tgctgccgac cgctgctgct ggtctgctgc
       61 tcctcgctgc ccagccggcc atggccaagc ttatcgatga taagctgtca aacatgagca
      121 gatctgagcc cgcctaatga gcgggctttt ttttcagatc tgcttgaaga cgaaagggcc
      181 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgatgc
      241 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
      301 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
      361 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
      421 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
      481 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
      541 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
      601 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
      661 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg
      721 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      781 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      841 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      901 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      961 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1021 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1081 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1141 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1201 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1261 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1321 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1381 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1441 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1501 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1561 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1621 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1681 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1741 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1981 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2341 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2401 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2461 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2521 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2581 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2641 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     2701 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     2761 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     2821 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2881 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagaacgcca
     2941 tcaaaaataa ttcgcgtctg gccttcctgt agccagcttt catcaacatt aaatgtgagc
     3001 gagtaacaac ccgtcggatt ctccgtggga acaaacggcg gattgaccgt aatgggatag
     3061 gttacgttgg tgtagatggg cgcatcgtaa ccgtgcatct gccagtttga ggggacgacg
     3121 acagtatcgg cctcaggaag atcgcactcc agccagcttt ccggcaccgc ttctggtgcc
     3181 ggaaaccagg caaagcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     3241 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     3301 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatcc
     3361 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3421 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     3601 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     3661 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     3721 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     3781 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     3841 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     3901 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     3961 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     4021 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     4081 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     4141 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     4201 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     4261 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     4321 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     4381 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     4441 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     4501 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     4561 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     4621 ccataccgcg aaaggttttg caccattcga tggtgtcctg gcacgacagg tttcccgact
     4681 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc
     4741 aggctttaca ctttatgctt ccggctcgta taatgtgtgg aattgtgagc ggataacaat
     4801 ttcacacagg aaacaggatc gatccatcga tgagcttact ccccatcccc ctgttgacaa
     4861 ttaatcatcg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     4921 caggatcagc ttactcccca tccccctgtt gacaattaat catcggctcg tataatgtgt
     4981 ggaattgtga gcggataaca atttcacaca ggaaacagga tcc
//
