LOCUS       Exported                5977 bp ds-DNA     circular SYN 15-MAR-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 15, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5977
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        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"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /note="color: #cc99b2
                     Cleavage site after base 935"
                     /translation="LEVLFQGP"
     misc_feature    951..1946
                     /locus_tag="p37 human"
                     /label=p37 human codon optimized for E.coli
                     /label=p37 human
                     /note="color: #808000"
     CDS             2385..3245
                     /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: 2385 .. 2453 / #ccffcc / signal sequence
                        2: 2454 .. 3245 / #ccffcc
                     Cleavage site after base 2453"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3416..4004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     promoter        4248..4325
                     /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             4326..5408
                     /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"
     protein_bind    5421..5442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        5457..5487
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 5457 .. 5462 / #ffffff / -35
                        2: 5463 .. 5480 / #ffffff
                        3: 5481 .. 5487 / #ffffff / -10"
     protein_bind    5495..5511
                     /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"
     primer_bind     5519..5535
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     CDS             join(5531..5977,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /note="color: #993366"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5551..5567)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
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 atggcggagg
      961 gcggcggtcc agaacctggt gaacaagagc gccgtagctc tggcccgcgc ccgccaagcg
     1021 cccgtgatct gcaattggca ctggcggagc tgtacgagga tgaggtgaaa tgcaaaagct
     1081 ccaaaagcaa ccgccctaag gcgacggtat ttaagtctcc tcgcacgccg ccgcaacgtt
     1141 tctatagcag cgagcacgag tatagcggcc ttaacatcgt ccgtccgtct accggcaaaa
     1201 tcgttaatga actgttcaaa gaagctcgcg aacacggtgc agttccactg aacgaagcaa
     1261 cgcgtgcgtc tggcgatgac aaaagcaaga gctttaccgg tggcggttac cgtttgggca
     1321 gcagcttttg taaacgttcc gaatatattt atggtgagaa ccagcttcag gacgttcaga
     1381 tccttcttaa actttggagc aacggcttta gcttggatga tggcgaactt cgcccttata
     1441 acgagccgac gaacgctcaa ttcctggaat ccgtcaaacg cggtgagatc ccacttgaac
     1501 tgcaacgcct ggttcacggc ggccaggtaa atcttgacat ggaagaccac caggatcaag
     1561 agtacatcaa gccgcgcctg cgctttaagg cctttagcgg cgagggtcag aagctgggct
     1621 ccctgacgcc tgaaatcgtg agcaccccta gcagcccgga agaggaagat aagtccattt
     1681 tgaatgcggt agtcttgatt gatgatagcg tcccgaccac caagattcaa atccgtctgg
     1741 cggatggcag ccgtcttatt caacgtttta acagcaccca tcgtatcctg gatgtccgca
     1801 atttcatcgt ccaaagccgc ccggaatttg ctgccctgga ctttattctt gtcacctctt
     1861 tcccgaataa agaacttacc gatgagagcc tgacccttct ggaagccgat attttgaata
     1921 ctgtgctgct gcaacaactg aaataagaat tcccgggtcg actcgagcgg ccgcatcgtg
     1981 actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2041 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2101 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     2161 atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     2221 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2281 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2341 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     2641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2761 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     2941 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3061 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3121 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3181 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3241 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3301 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3361 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3421 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3481 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3541 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3601 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3661 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3721 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3781 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3841 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3901 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3961 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4021 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4081 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4141 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4201 attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat
     4261 ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     4321 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     4381 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     4441 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     4501 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     4561 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     4621 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     4681 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     4741 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     4801 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     4861 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     4921 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     4981 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     5041 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     5101 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     5161 gctcatgtta tatcccgccg tcaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     5221 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     5281 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     5341 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     5401 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     5461 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     5521 ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg
     5581 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5641 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5701 cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg
     5761 cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     5821 tgcgcccatc tacaccaacg taacctatcc cattacggtc aatccgccgt ttgttcccac
     5881 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     5941 aggccagacg cgaattattt ttgatggcgt tggaatt
//
