LOCUS       62056_11500        4994 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 4994)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4994
                     /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             289..306
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             328..981
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     promoter        1297..1401
                     /label=AmpR promoter
     CDS             1402..2259
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2433..3021
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        3265..3342
                     /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             3343..4422
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    4438..4459
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4474..4504
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     CDS             4548..4721
                     /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 agtattcgga tccccggaat tcccgggtcg acctcgagct ggtgccgcgc
      301 ggcagctcag gaggaagcgg agggagtatg tcccctatac taggttattg gaaaattaag
      361 ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat
      421 ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag
      481 tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc
      541 atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag
      601 atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat
      661 agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa
      721 atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct
      781 gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat
      841 gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag
      901 tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt
      961 ggtggcgacc atcctccaaa atagcggccg catcgtgact gactgacgat ctgcctcgcg
     1021 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     1081 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1141 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga
     1201 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1261 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1321 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1381 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1441 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1501 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1561 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1621 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     1681 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1741 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1801 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1861 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1921 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     1981 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2041 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2101 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2161 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2221 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2281 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2341 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2401 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2461 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2521 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2581 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2641 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2701 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2761 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2821 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2881 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2941 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3001 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3061 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3121 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3181 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     3241 ggtatttcac accgcataaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg
     3301 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     3361 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     3421 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     3481 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     3541 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     3601 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     3661 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     3721 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     3781 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     3841 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     3901 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     3961 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     4021 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     4081 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     4141 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     4201 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     4261 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     4321 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4381 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4441 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4501 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     4561 cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     4621 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     4681 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc
     4741 cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg
     4801 tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa
     4861 cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact
     4921 cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg
     4981 atggcgttgg aatt
//
