LOCUS       62056_20905        7969 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 7969)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             918..941
                     /codon_start=1
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
                     /translation="LEVLFQGP"
     promoter        4317..4421
                     /label=AmpR promoter
     CDS             4422..5234
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5408..5996
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6240..6317
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6318..7397
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    7413..7434
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7449..7479
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             7523..7696
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc atgaccatga
      961 ttacgccaag cttagccgcc agcgaggcca cgggcggcct ggatcgcgaa cgcatcgact
     1021 tgttgtggaa aatcgctcgc gccggcgccc gttccgcagt gggcaccgag gcgcgtcgcc
     1081 agttccgcta cgacggcgac atgaatatcg gcgtgatcac cgatttcgag ctggaagtgc
     1141 gcaatgcgct gaacaggcgg gcgcacgccg tcggcgcgca ggacgtggtc cagcatggca
     1201 ctgagcagaa caatcctttc ccggaggcag atgagaagat tttcgtcgta tcggccaccg
     1261 gtgaaagcca gatgctcacg cgcgggcaac tgaaggaata cattggccag cagcgcggcg
     1321 agggctatgt cttctacgag aaccgtgcat acggcgtggc ggggaaaagc ctgttcgacg
     1381 atgggctggg agccgcgccc ggcgtgccga gcggacgttc gaagttctcg ccggatgtac
     1441 tggaaacggt gccggcgtca cccggattgc ggcggccgtc gctgggcgca gtggaacgcc
     1501 actgcaggtc gactctagag gatcccatgc gcctgctgaa gttcctgtgg tggacttgcg
     1561 tcaccctgat ttgtggagtg ctgctcagtt tcagcggcgc ctatctctat ctgagtccca
     1621 gcctgccctc ggtggaagct ctgcgtaacg tccagctgca gatccccctc aaggtctaca
     1681 gcgaggacgg caagctgatc tccgagttcg gcgagatgcg ccggacgccg atccgcttcg
     1741 ccgacatccc gcaggacttc atccatgccc tgctgtccgc cgaggacgac aatttcgcca
     1801 accattatgg cgtcgatgtg aaaagcctaa tgcgcgccgc cgcgcaatta ttgaaaagcg
     1861 gtcacatcca gactggcggc agcaccatca ccatgcaggt agcgaagaac tacttcctta
     1921 ccaatgaaag aagcttctcg cgaaagatca atgaaatcct cctggccctg cagatcgagc
     1981 gacagctgac caaggacgag atcctcgagc tctacgtcaa caagatctac ctgggcaacc
     2041 gtgcctatgg catcgaggct gccgcacagg tctattacgg caagccgatc aaagacttga
     2101 gcctggccga gatggcgatg atcgccggcc tgccgaaagc gccctcgcgc tacaacccgc
     2161 tggtcaaccc gacccgcagt acggagcggc gcaactggat cctcgagcgc atgctcaagc
     2221 tcggcttcat cgaccagcaa cgctaccagg ccgccgtcga ggagccgatc aacgcctcct
     2281 accacgtgca gacccccgaa ctgaatgcgc cctacatcgc cgagatggcc cgtgccgaga
     2341 tggtcggccg ctacggcagc gaggcctata ccgagggcta caaggtcatc accacggtcc
     2401 gcagcgacct gcagaacgcc gccagccaat cggtccgcga cggcctcatc gactatgacc
     2461 agcgccacgg ctaccgcggc ccggagaccc gcctgccggg ccagacccgc gacgcctggc
     2521 tcaagcacct gggccagcag cgcagcatcg gcggcctgga gccggcgatc gtcacccagg
     2581 tggaaaagag cgggatcatg gtgatgaccc gcgacggcaa ggaagaagcc gtgacctggg
     2641 acagcatgaa atgggcccgc cccttcctca gcaacaacag catgggcccg atgccgcgcc
     2701 agcccgccga cgtggcccag gccggcgacc agatccgcgt gcagcgccag gaagacggca
     2761 ccctgcgctt cgtacagact cccgccgcac agagcgcgct gatctccctg gacccgaagg
     2821 acggggcgat ccgctcgctg gtcggcggct tctccttcga gcagagcaac tacaaccgcg
     2881 cgatccaggc caagcgccag ccgggctcca gtttcaagcc gttcatctac agcgcggccc
     2941 tggacaacgg cttcaccgcc gccagcctgg tcaacgatgc gccgatcgtg ttcgtcgacg
     3001 agtacctgga caaggtctgg cgaccgaaga acgacaccaa taccttcctc ggcccgatcc
     3061 cgctgcgcga agccctgtac aagtcccgca acatggtctc gatccgcgtc ctgcagggcc
     3121 tgggcatcga gcgggcgatc agctacatca ccaagttcgg cttccagcgc gacgaactgc
     3181 cgcgcaactt ctccctggcc ctgggtaccg cgacagtcac accgatggaa atcgccggcg
     3241 cctggagcgt attcgccaac ggcggctaca aggtcaatcc gtacgtcatc gaacgcatcg
     3301 agagccgcga cggacaggtg ctgtaccagg ccaatccgcc ccgcgtgccg gtggaagaac
     3361 aggtcgctgc gccggatgcg gaagacgccg ggaaacctgg cgaccccgag catccggaga
     3421 gcgccgaagg cgaaggttcg atcgaagccc agcaagtcgc cgccaaggcc cagaccacct
     3481 tcgagccgac cccggccgaa cggatcatcg atgcccgtac cgcctatatc atgaccagca
     3541 tgctgcagga cgtgatcaag cgcggtaccg ggcgccgcgc cctggcgctg aagcgcaccg
     3601 acctggccgg caagaccggc accaccaacg attccaagga tggctggttc tccggctaca
     3661 actccgacta cgtgaccagc gtctgggtcg gcttcgacca gccggagacc ctgggtcgcc
     3721 gcgagtacgg cggtaccgtc gcgctgccga tctggatcag gtacatgggc ttcgccctca
     3781 aggacaagcc gatgcacacc atggccgagc cgccaggcat cgtcagcctg cgcatcgacc
     3841 cggtaaccgg ccgttcggcc gcgccgggca ctcccggcgc ctatttcgag atgttcaaga
     3901 acgaggacac cccgccctcg gtcaacgaac tgccgccggg cagcttccct ggcagcccgc
     3961 tgccggatga cgaaggcgcg ccgatcgacc tgttctgact cgagcggccg catcgtgact
     4021 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4081 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4141 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     4201 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4261 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4321 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4381 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagccat attcaacggg
     4441 aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     4501 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     4561 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     4621 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     4681 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat
     4741 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     4801 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     4861 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     4921 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     4981 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     5041 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     5101 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     5161 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     5221 tcgatgagtt tttctaactg tcagaccaag tttactcata tatactttag attgatttaa
     5281 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5341 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5401 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5461 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5521 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     5581 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5641 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5701 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5761 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5821 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5881 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5941 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6001 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6061 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6121 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
     6241 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     6301 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     6361 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     6421 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     6481 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     6541 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     6601 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     6661 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     6721 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     6781 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     6841 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     6901 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     6961 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     7021 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     7081 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     7141 ataccgaaga cagctcatgt tatatcccgc cgtcaaccac catcaaacag gattttcgcc
     7201 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     7261 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     7321 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     7381 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7441 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7501 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
     7561 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     7621 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     7681 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
     7741 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
     7801 gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc
     7861 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     7921 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt
//
