LOCUS       62056_7810        5079 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 5079)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        86..163
                     /label=lacI promoter
     CDS             164..1243
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    1259..1280
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1418..2006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2180..3037)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3038..3142)
                     /label=AmpR promoter
     rep_origin      3242..3697
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     3'UTR           complement(3896..4040)
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     CDS             4054..4071
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(4129..4146)
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             complement(4153..4806)
                     /codon_start=1
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             complement(4810..4824)
                     /codon_start=1
                     /label=TEE
                     /note="translation enhancing element for E. coli (Qing et
                     al., 2004)"
                     /translation="MNHKV"
     5'UTR           complement(4825..4958)
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
     protein_bind    complement(4980..4996)
                     /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)."
     promoter        complement(4999..5065)
                     /label=cspA promoter
                     /note="promoter of the E. coli cold shock protein cspA gene
                     (Mitta et al., 1997)"
ORIGIN
        1 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
       61 cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat
      121 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
      181 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
      241 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
      301 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
      361 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
      421 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
      481 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
      541 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
      601 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
      661 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
      721 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
      781 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
      841 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
      901 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
      961 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     1021 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     1081 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     1141 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1201 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     1261 atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca
     1321 acccagtcag ctccttccgg tgggcgcggg gcatgactat gtgagcaaaa ggccagcaaa
     1381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1801 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2221 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2281 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     2341 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     2401 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     2461 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     2521 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     2581 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     2641 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     2701 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     2761 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     2821 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     2881 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     2941 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3001 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3061 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3121 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcctgatgcg
     3181 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     3241 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3421 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     3481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3541 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
     3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3661 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     3721 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3781 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     3841 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctgcgc
     3901 attctcattg cacccaaatt tattcttcac aaaaataata atagatttta ttacgcgatc
     3961 gattatttat ttcctgaaaa caaataaaaa aatccccgcc aaatggcagg gatcttagat
     4021 tctgtgcttt taagcagaga ttacctatct agacatcatc atcatcatca tctgcaggtc
     4081 gacaagcttg aattcggatc cctcgagggt accgagctcc atatgtccgc tgccccgggg
     4141 gaccagatcc gattttggag gatggtcgcc accaccaaac gtggcttgcc agccctgcaa
     4201 aggccatgct atatacttgc tggatttcaa gtacttatca atttgtggga tagcttcaat
     4261 acgtttttta aaacaaacta attttgggaa cgcatccagg cacattgggt ccatgtataa
     4321 aacaacatca agagcgtcat acaacatgaa gtcaggatgg gttacatgat caccatttaa
     4381 atatgtttta tgacataaac gatcttcgaa cattttcagc atttcaggta gcttgctaag
     4441 aaaatcaact ttgagagttt caaagtcttt actatatgca attctcgaaa caccgtatct
     4501 aatatccaaa accgctcctt caagcattga aatctctgca cgctcttttg gacaaccacc
     4561 caacatgttg tgcttgtcag ctatataacg tatgatggcc atagactgtg ttaatttaac
     4621 atcaccatca atataataag gaagattggg aaactccaaa cccaattcaa actttttgtt
     4681 tcgccattta tcaccttcat cgcgctcata caaatgctct tcatattttt cttcaagata
     4741 ttccaaaaga agtcgagtgg gttgcacaag gcccttaatt ttccaataac ctagtatagg
     4801 ggacatgtgc actttgtgat tcatggtgta ttacctctta ataattaagt gtgcctttcg
     4861 gcgatatggc gtgctttaca gattttgaag cgttaaagga atgtgcacta cgaggggtat
     4921 caacgataac tcttgaaggg acttgcctta ctacactgga tatgcgctag cacatcaaat
     4981 tgttatccgc tcacaatttg atgtgcatta agccacgcat tggcgggtga tgcaacaatt
     5041 atttttcata tttatgatta atcggccaca ccattcctt
//
