LOCUS       Exported                7067 bp ds-DNA     circular SYN 30-NOV-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    GST-mDia1 (FH1-FH2-DAD)-His
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7067)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, December 22, 2016 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7067
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    183..211
                     /note="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /note="GST"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /note="HRV 3C site"
                     /translation="LEVLFQGP"
     CDS             948..3011
                     /codon_start=1
                     /note="mDia1 558-1255"
                     /translation="SAAVPPAPPLPGDSGTVIPPPPPPPPLPGGVVPPSPPLPPGTCIP
                     PPPPLPGGACIPPPPQLPGSAAIPPPPPLPGVASIPPPPPLPGATAIPPPPPLPGATAI
                     PPPPPLPGGTGIPPPPPPLPGSVGVPPPPPLPGGPGLPPPPPPFPGAPGIPPPPPGMGV
                     PPPPPFGFGVPAAPVLPFGLTPKKVYKPEVQLRRPNWSKFVAEDLSQDCFWTKVKEDRF
                     ENNELFAKLTLAFSAQTKTSKAKKDQEGGEEKKSVQKKKVKELKVLDSKTAQNLSIFLG
                     SFRMPYQEIKNVILEVNEAVLTESMIQNLIKQMPEPEQLKMLSELKEEYDDLAESEQFG
                     VVMGTVPRLRPRLNAILFKLQFSEQVENIKPEIVSVTAACEELRKSENFSSLLELTLLV
                     GNYMNAGSRNAGAFGFNISFLCKLRDTKSADQKMTLLHFLAELCENDHPEVLKFPDELA
                     HVEKASRVSAENLQKSLDQMKKQIADVERDVQNFPAATDEKDKFVEKMTSFVKDAQEQY
                     NKLRMMHSNMETLYKELGDYFVFDPKKLSVEEFFMDLHNFRNMFLQAVKENQKRRETEE
                     KMRRAKLAKEKAEKERLEKQQKREQLIDMNAEGDETGVMDSLLEALQSGAAFRRKRGPR
                     QVNRKAGCAVTSLLASELTKDDAMAPGPVKVPKKSEGVPTILEEAKELVGRAS"
     misc_feature    1002..1487
                     /note="FH1"
     misc_feature    1500..2708
                     /note="FH2 Domain"
     misc_feature    2775..2861
                     /note="DAD Domain"
     CDS             3018..3041
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /note="8xHis"
                     /translation="HHHHHHHH"
     promoter        3370..3474
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             3475..4335
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4506..5094
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5338..5415
                     /gene="lacI (mutant)"
                     /note="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5416..6498
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    6511..6532
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6547..6577
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6585..6601
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     primer_bind     6609..6625
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6621..81
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6641..6657)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 gctgctgttc
      961 cccctgcccc tcctctgcct ggtgactctg gcactgttat tccacctccc ccacccccac
     1021 ctcctcttcc tggaggtgtg gtcccaccat cccctcctct gcctccaggt acttgtatcc
     1081 ctccacctcc tcctttacct ggaggtgctt gtataccccc tcccccccag ttgcctggca
     1141 gtgctgccat ccctccacct cctcctctac ctggagttgc ttccatcccc ccacctcccc
     1201 ctttgcctgg ggctactgcc atccccccac ctcccccttt gcctggggct actgccatcc
     1261 ccccacctcc ccctttgcct ggaggtacag gtataccacc accacctcct cctttgcctg
     1321 gaagtgttgg cgttccccca ccccctccct tgcctggagg accaggactg cctcctcccc
     1381 ctcccccttt tcctggagca cctggcattc ctccacctcc acctggtatg ggcgtgcctc
     1441 cacctccccc ctttggattt ggggttcctg cggccccagt tctgccattt ggattaaccc
     1501 ccaaaaaagt ttataagcca gaggtgcagc tccggaggcc aaactggtcc aagtttgtgg
     1561 ctgaggacct ttcccaggac tgcttctgga caaaggtgaa ggaggaccgc tttgagaaca
     1621 atgaactttt tgccaaactt acccttgcct tctccgccca gaccaagact tctaaagcca
     1681 agaaggatca agaaggtgga gaagaaaaga aatctgttca aaagaagaaa gtaaaagagc
     1741 tgaaagtgct ggattcaaag acagcgcaga atctctcaat ctttttgggt tcattccgca
     1801 tgccctatca agagataaag aacgttatcc tggaggtgaa tgaggctgtt ctcacagagt
     1861 ctatgatcca gaacctcatt aaacagatgc cagagccaga gcagctaaag atgctctctg
     1921 aactgaagga ggagtacgat gatctggctg agtcagagca gtttggtgtg gtgatgggca
     1981 cagtgccccg ccttcggcct cgcctcaacg ccatcctctt caagctacag ttcagtgagc
     2041 aagttgagaa catcaagcca gagatcgtgt ctgtcaccgc cgcatgcgaa gagctgcgta
     2101 agagtgagaa cttctccagc ctcctggagc tcacactgct ggtcggaaac tatatgaatg
     2161 cgggctccag gaatgctggt gctttcggct tcaatatcag cttcctttgt aagcttcgag
     2221 acaccaagtc tgcagatcag aagatgactc tgttgcattt cttggctgag ttatgtgaga
     2281 atgaccaccc cgaagtcctc aagtttcctg atgagcttgc ccatgtagag aaagccagca
     2341 gagtctctgc tgagaacctg cagaagagct tagatcagat gaagaagcag attgcggacg
     2401 tggagcgcga tgttcagaat ttcccagctg ccactgacga gaaggacaag tttgttgaga
     2461 agatgaccag ctttgtgaag gatgcacagg aacagtataa caaactacgg atgatgcact
     2521 ccaacatgga gaccctctat aaggagctag gtgactactt cgtctttgac cctaagaagt
     2581 tgtctgtaga ggaattcttt atggatctgc acaactttag gaatatgttt ttgcaagcag
     2641 tcaaggaaaa ccagaagcgc cgggaaacag aagaaaagat gcggagagca aaattagcca
     2701 aggagaaggc agaaaaagag cgactggaga agcagcagaa gcgcgagcag ctcatcgaca
     2761 tgaacgcaga gggggatgag acaggtgtga tggacagtct tctagaagct ctgcagtcag
     2821 gggcagcatt ccgacggaag agagggcccc ggcaggtcaa caggaaggct gggtgtgcag
     2881 tcacatctct gctagcctcg gagctgacca aggatgatgc catggctcct ggtcctgtta
     2941 aggtacccaa gaaaagtgaa ggagtcccca caatcctgga agaagccaag gagctggttg
     3001 gccgtgcaag cctcgagcac caccaccacc accaccacca ctaattgatt aataccgcgg
     3061 ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     3121 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     3181 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     3241 tcacgtagcg atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta
     3301 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3361 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3421 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3481 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3541 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3601 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3661 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     3721 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3781 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3841 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3901 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3961 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     4021 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4081 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4141 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4201 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4261 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4321 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4381 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4441 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4501 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4561 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4621 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4681 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4741 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4801 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4861 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4921 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4981 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5041 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5101 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5161 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5221 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5281 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac
     5341 accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     5401 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     5461 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     5521 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     5581 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     5641 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     5701 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     5761 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     5821 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     5881 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     5941 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     6001 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     6061 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     6121 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     6181 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     6241 accgaagaca gctcatgtta tatcccgccg tcaaccacca tcaaacagga ttttcgcctg
     6301 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     6361 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     6421 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     6481 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     6541 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     6601 atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac
     6661 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6721 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6781 gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct
     6841 ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc
     6901 acggttacga tgcgcccatc tacaccaacg taacctatcc cattacggtc aatccgccgt
     6961 ttgttcccac ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct
     7021 ggctacagga aggccagacg cgaattattt ttgatggcgt tggaatt
//
