LOCUS       62056_20915        6472 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 6472)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6472
                     /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..935
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     promoter        2775..2879
                     /label=AmpR promoter
     CDS             2880..3737
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3911..4499
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4743..4820
                     /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             4821..5900
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    5916..5937
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5952..5982
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             6026..6199
                     /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 gttccgcgtg gatccccgga attcatgatt gacccatctg
      961 ccacttcccg gtatggttcc ccacgtttag ttagtaatgg ttttcggcat cggagaaaag
     1021 tggtgtatca gcgggtaggg cacaggcgat tttctctcat tttctttttc gttgtggttc
     1081 tggggcggtc cccgcggctg tgggctcagg tttcgttcac cccggatatt gaaggctatg
     1141 cggagctggc ctggggcatt gcatccgatc gtggcgccct caagcatgga tttaagacta
     1201 ctactgattt taagattgtg ttccccattg tggcaaagaa ggatttcaag taccgcggtg
     1261 aggggaatgt ctatgcggaa attaatgtta aagcgttgaa gttgagttta gagtcaaatg
     1321 gtggagcaaa gtttgacacg aagggttctg caaagacgat agaggcaacc ctgcactgtt
     1381 atggggccta cctgaccatt gggaagaatc ctgattttaa gtcaacgttt gctgttttgt
     1441 gggagccgtg gaccgcgaat ggggattata agtctaaggg agataagccg gtgtatgagc
     1501 cggggtttga gggagccggg ggaaagttag ggtataaaca gactgacatc gccggcacgg
     1561 ggctcacgtt tgatattgcg tttaagtttg cgtctaacac cgactgggag ggcaaagaca
     1621 gcaagggcaa cgtcccagca ggagtaaccc ccagcaagta tggattgggg ggagatattt
     1681 tgttcggctg ggagcgtacg cgtgaagatg gcgtgcagga atacattaaa gtggagctca
     1741 ccggcaactc cacactgtct agcgactatg cccaagcccg agccccggca gccggggcta
     1801 aggtgagtat gaagctttgg ggtctgtgtg ctctggctgc tacagacgtg gggcataaga
     1861 aaaacggagc gcagggcacc gtaggcgcag atgcgttgtt gacgttgggg tatcgttggt
     1921 tctcggcggg aggatatttc gcatcgcagg ccagcaatgt attcggggga gtatttctca
     1981 acatggccat gcgagagcac gactgtgctg cctatattaa gctcgaaacc aaggggtctg
     2041 atcctgatac ttctttcctt gagggtcttg atttgggtgt tgatgtgcgt acgtacatgc
     2101 ctgtccatta caaagtccta aaagccctac cccgagccga catccacttc ccggtgtatg
     2161 gaaaagtctg gggttcgtat cgtcatgata tgggtgagta tggttgggtt aaagtgtatg
     2221 caaacttgta cggcggtacg aacaaaaagg ccacgccccc tgctgctcct gctacgaagt
     2281 ggagcaagga atattgtggg tattacgagt gtggggtagt ggtcagtccg ttagagaagg
     2341 tggagattcg gctgagctgg gagcaaggca agctacaaga gaacagcaat gtagtgatag
     2401 agaagaacgt gacggagcgt tggcaattcg taggggcatg tcgcttgatt tggtagctcg
     2461 agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa
     2521 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2581 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     2641 cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac
     2701 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2761 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2821 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2881 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2941 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3001 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3061 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3121 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3181 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3241 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3301 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3361 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3421 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3481 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3541 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3601 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3661 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3721 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3781 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3841 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3901 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3961 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4021 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4081 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4141 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4201 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4261 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4321 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4381 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4441 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4501 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4561 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4621 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4681 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
     4741 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     4801 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     4861 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     4921 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     4981 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     5041 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     5101 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     5161 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     5221 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     5281 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     5341 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     5401 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     5461 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     5521 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     5581 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     5641 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     5701 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     5761 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     5821 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     5881 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     5941 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6001 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     6061 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     6121 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     6181 gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag cggtgccgga
     6241 aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct caaactggca
     6301 gatgcacggt tacgatgcgc ccatctacac caacgtaacc tatcccatta cggtcaatcc
     6361 gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta atgttgatga
     6421 aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa tt
//
