LOCUS       62056_9505        5632 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 5632)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             89..121
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     terminator      189..236
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(411..439)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        552..656
                     /label=AmpR promoter
     CDS             657..1469
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1643..2231
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2417..2559)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2664..2852)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(4130..4151)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4167..5246)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5247..5324)
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat ggctagcatg actggtggac agcaaatggg
      121 tcgcggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      181 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
      241 aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
      301 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
      361 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt
      421 atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc
      481 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      541 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      601 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      661 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
      721 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
      781 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
      841 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
      901 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
      961 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1021 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1081 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1141 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     1201 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     1261 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     1321 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     1381 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     1441 agtttcattt gatgctcgat gagtttttct aactgtcaga ccaagtttac tcatatatac
     1501 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2281 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2341 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2401 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2461 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2521 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2581 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2641 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2701 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2761 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2821 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2881 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     2941 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3001 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     3061 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3121 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3181 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3241 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3301 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     3361 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     3421 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     3481 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     3541 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     3601 cggcataaat cgccgtgacg atcagcggtc cagtgatcga agttaggctg gtaagagccg
     3661 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     3721 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     3781 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     3841 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     3901 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     3961 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4021 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     4081 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     4141 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     4261 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     4321 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     4381 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     4441 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     4501 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     4561 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     4621 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     4681 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     4741 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     4801 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     4861 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     4921 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     4981 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     5041 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     5101 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     5161 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     5221 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     5281 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     5341 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     5401 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     5461 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     5521 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     5581 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at
//
