LOCUS       Exported                6857 bp DNA     circular SYN 15-MAR-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6857)
  AUTHORS   gene
  TITLE     Direct Submission
  JOURNAL   Exported Jun 14, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
                     /note="This forward directional feature has 3 segments:
                        1: 183 .. 188 / #ffffff / -35
                        2: 189 .. 204 / #ffffff
                        3: 205 .. 211 / #ffffff / -10"
     protein_bind    219..235
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
                     /note="color: #31849b"
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /note="color: #cc99b2"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5' Sequencing Primer
                     /note="color: black; sequence: GGGCTGGCAAGCCACGTTTGGTG"
     CDS             918..939
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=PreScission site
                     /note="color: #cc99b2
                     Cleavage site after base 935"
                     /translation="LEVLFQG"
     misc_feature    2859..2869
                     /label=stop codons
                     /note="stop codons in all three reading frames"
                     /note="color: #ff0000"
     primer_bind     complement(2907..2929)
                     /label=pGEX 3' Sequencing Primer
                     /note="color: black; sequence: CCGGGAGCTGCATGTGTCAGAGG"
     promoter        3160..3264
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3265..4125
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3265 .. 3333 / #ccffcc / signal sequence
                        2: 3334 .. 4125 / #ccffcc
                     Cleavage site after base 3333"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4296..4884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     promoter        5128..5205
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5206..6288
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
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 cctgggagta cgcgccggct
      961 ccgaacaggg ccatctacca ccgcccagtg ccccagctgg gagagcgcgt tcatcaactg
     1021 tcacgggtgg tgaggaacca acgccatcag tggcctatgt acatactacg ccgggtctgc
     1081 cttcaggctg ggaagaaaga aaagatgcta aggggcgcac atactatgtc aatcataaca
     1141 atcgaaccac aacttggact cgacctatca tgcagcttgc agaagatggt gcgtccggat
     1201 cagccacaaa cagtaacaac catctaatcg agcctcagat ccgccggcct cgtagcctca
     1261 gctcgccaac agtaacttta tctgccccgc tggagggtgc caaggactca cccgtacgtc
     1321 gggctgtgaa agacaccctt tccaacccac agtccccaca gccatcacct tacaactccc
     1381 ccaaaccaca acacaaagtc acacagagct tcttgccacc cggctgggaa atgaggatag
     1441 cgccaaacgg ccggcccttc ttcattgatc ataacacaaa gactacaacc tgggaagatc
     1501 cacgtttgaa atttccagta catatgcggt caaagacatc tttaaacccc aatgaccttg
     1561 gcccccttcc tcctggctgg gaagaaagaa ttcacttgga tggccgaacg ttttatattg
     1621 atcataatag caaaattact cagtgggaag acccaagact gcagaaccca gctattactg
     1681 gtccggctgt cccttactcc agagaattta agcagaaata tgactacttc aggaagaaat
     1741 taaagaaacc tgctgatatc cccaataggt ttgaaatgaa acttcacaga aataacatat
     1801 ttgaagagtc ctatcggaga attatgtccg tgaaaagacc agatgtccta aaagctagac
     1861 tgtggattga gtttgaatca gagaaaggtc ttgactatgg gggtgtggcc agagaatggt
     1921 tcttcttact gtccaaagag atgttcaacc cctactacgg cctctttgag tactctgcca
     1981 cggacaacta cacccttcag atcaacccta attcaggcct ctgtaatgag gatcatttgt
     2041 cctacttcac ttttattgga agagttgctg gtctggccgt atttcatggg aagctcttag
     2101 atggtttctt cattagacca ttttacaaga tgatgttggg aaagcagata accctgaatg
     2161 acatggaatc tgtggatagt gaatattaca actctttgaa atggatcctg gagaatgacc
     2221 ctactgagct ggacctcatg ttctgcatag acgaagaaaa ctttggacag acatatcaag
     2281 tggatttgaa gcccaatggg tcagaaataa tggtcacaaa tgaaaacaaa agggaatata
     2341 tcgacttagt catccagtgg agatttgtga acagggtcca gaagcagatg aacgccttct
     2401 tggagggatt cacagaacta cttcctattg atttgattaa aatttttgat gaaaatgagc
     2461 tggagttgct catgtgcggc ctcggtgatg tggatgtgaa tgactggaga cagcattcta
     2521 tttacaagaa cggctactgc ccaaaccacc ccgtcattca gtggttctgg aaggctgtgc
     2581 tactcatgga cgccgaaaag cgtatccggt tactgcagtt tgtcacaggg acatcgcgag
     2641 tacctatgaa tggatttgcc gaactttatg gttccaatgg tcctcagcta tttacaatag
     2701 agcaatgggg cagtcctgag aaactgccca gagctcacac atgctttaat cgccttgact
     2761 tacctccata tgaaaccttt gaagatttac gagagaaact tctcatggcc gtggaaaatg
     2821 ctcaaggatt tgaaggggtg gattaagcgg ccgcatcgtg actgactgac gatctgcctc
     2881 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     2941 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     3001 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct
     3061 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3121 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3181 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3241 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3301 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3361 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3421 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3481 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     3541 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3601 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3661 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3721 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3781 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     3841 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3901 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3961 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4021 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4081 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4141 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4201 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4441 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5041 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     5101 tgcggtattt cacaccgcat aaattccgac accatcgaat ggtgcaaaac ctttcgcggt
     5161 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     5221 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     5281 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     5341 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     5401 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     5461 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     5521 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     5581 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     5641 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     5701 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     5761 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     5821 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     5881 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     5941 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     6001 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     6061 tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     6121 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     6181 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6241 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6301 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     6361 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     6421 ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6481 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6541 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt
     6601 ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata
     6661 ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg
     6721 taacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt
     6781 actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt
     6841 ttgatggcgt tggaatt
//
