LOCUS       62056_10410        5546 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 5546)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     155..171
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        207..225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    226..250
                     /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             254..276
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             287..304
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             311..964
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             986..1006
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     promoter        1743..1820
                     /label=lacI promoter
     CDS             1821..2900
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    2916..2937
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2952..2982
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     3014..3030
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3046..3062)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3354..3942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4116..4973)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4974..5078)
                     /label=AmpR promoter
     rep_origin      5105..5546
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
       61 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
      121 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccgaattc
      181 tgctggactt tccaaaatgt cgagcttaat acgactcact ataggggaat tgtgagcgga
      241 taacaattcc ccttttgttt aactttaaga aggagaggcc accatgcatc accatcacca
      301 tcacgcaggc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
      361 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
      421 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
      481 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
      541 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
      601 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
      661 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
      721 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
      781 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
      841 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
      901 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
      961 aaaatcggat atcatcccaa cgaccgaaaa cctgtatttt cagggcgcga tcgccggcgc
     1021 gccagatctc aagcttggcg gcggagggag tggcggcgga gggagtatga tggacttgga
     1081 gctgccgccg ccgggactcc cgtcccagca ggacatggat ttgattgaca tactttggag
     1141 gcaagagata gatcttggag taagtcgaga agtatttgac ttcagtcagc gacggaaaga
     1201 gtatgagctg gaaaaacaga aaaaacttga aaaggaaaga caagaacaac tccaaaagga
     1261 gcaagagaaa gcctttttcg ctcagttaca actagatgaa gcagctgcag cttttctccc
     1321 aattcagcca gcccagcaca tccagtcaga aaccagttga gctagcggac cgacgcgtta
     1381 aacgcggccg ctcgaggttt aaacggccgg ccacatgact gactgagcca cgataaccgc
     1441 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttccc
     1501 accagccttg tcctaatccg cggcggttcc tggccttttg ctggcctttt gctcacatgt
     1561 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1621 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1681 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
     1741 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     1801 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     1861 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     1921 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     1981 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     2041 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     2101 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     2161 ttctcgcgca acgagtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     2221 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     2281 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     2341 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     2401 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     2461 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     2521 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     2581 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     2641 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     2701 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     2761 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     2821 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2881 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     2941 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     3001 taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg ccgtcgtttt
     3061 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     3121 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatggatccg ctcttccgct
     3181 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3241 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3301 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3361 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3421 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3481 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3541 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3601 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3661 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3721 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3781 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3841 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3901 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3961 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4021 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4081 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4141 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4201 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4261 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4321 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4381 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4441 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4501 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4561 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4621 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4681 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4741 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4801 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4861 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4921 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4981 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5041 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5101 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5521 tttaacgcga attttaacaa aatatt
//
