LOCUS       Exported                5810 bp ds-DNA     circular SYN 20-SEP-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGEX4T3-GST-NQO1(Y128F)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 27, 2021 from SnapGene 3.3.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5810
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        183..211
                     /note="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_feature    184..211
                     /label=tac promoter
                     /note="tac promoter"
                     /note="ApEinfo_fwdcolor=#00ff00
                     ApEinfo_revcolor=#00ff00"
     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)."
     misc_feature    258..935
                     /label=GST
                     /note="GST"
                     /note="ApEinfo_fwdcolor=#ff0000
                     ApEinfo_revcolor=#ff0000"
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /note="GST"
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     misc_feature    918..935
                     /label=Thrombin
                     /note="Thrombin"
                     /note="ApEinfo_fwdcolor=cyan
                     ApEinfo_revcolor=cyan"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     misc_feature    936..956
                     /label=Tev site
                     /note="Tev site"
                     /note="ApEinfo_fwdcolor=#ff8000
                     ApEinfo_revcolor=#ff8000"
     CDS             936..956
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /note="TEV site"
                     /translation="ENLYFQG"
     misc_feature    957
                     /label=New Feature(1)
                     /note="New Feature(1)"
                     /note="ApEinfo_fwdcolor=cyan
                     ApEinfo_label=New Feature
                     ApEinfo_revcolor=green"
     misc_feature    958..974
                     /label=GST(1)
                     /note="GST(1)"
                     /note="ApEinfo_fwdcolor=#ff0000
                     ApEinfo_label=GST
                     ApEinfo_revcolor=#ff0000"
     misc_feature    959..964
                     /label=EcoRI
                     /note="EcoRI"
                     /note="ApEinfo_fwdcolor=#00ff00
                     ApEinfo_revcolor=#00ff00"
     misc_feature    965..969
                     /label=SmaI
                     /note="SmaI"
                     /note="ApEinfo_fwdcolor=#ff8000
                     ApEinfo_revcolor=#ff8000"
     misc_feature    975..1799
                     /note="NQO1-Y128F"
     misc_feature    1359..1361
                     /note="Y128F"
     misc_feature    1800..1817
                     /label=GST(2)
                     /note="GST(2)"
                     /note="ApEinfo_fwdcolor=#ff0000
                     ApEinfo_label=GST
                     ApEinfo_revcolor=#ff0000"
     misc_feature    1860..1881
                     /label=pGEX 3 primer
                     /note="pGEX 3 primer"
                     /note="ApEinfo_fwdcolor=#00ff00
                     ApEinfo_revcolor=#00ff00"
     promoter        2113..2217
                     /gene="bla"
                     /note="AmpR promoter"
     misc_feature    2148..2175
                     /label=AmpR promoter
                     /note="AmpR promoter"
                     /note="ApEinfo_fwdcolor=#00ff00
                     ApEinfo_revcolor=#00ff00"
     misc_feature    2218..3078
                     /label=Ampicillin
                     /note="Ampicillin"
                     /note="ApEinfo_fwdcolor=#ff0000
                     ApEinfo_revcolor=#ff0000"
     CDS             2218..3078
                     /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      3249..3837
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4081..4158
                     /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."
     misc_feature    4150
                     /label=New Feature
                     /note="New Feature"
                     /note="ApEinfo_fwdcolor=#ff0000
                     ApEinfo_revcolor=#ff0000"
     misc_feature    4151..5241
                     /label=lacI
                     /note="lacI"
                     /note="ApEinfo_fwdcolor=#ff0000
                     ApEinfo_revcolor=#ff0000"
     CDS             4159..5241
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    5254..5275
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5290..5320
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    5328..5344
                     /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     5352..5368
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5364..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(5384..5400)
                     /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 gttccgcgtg gatccgaaaa cctgtatttt cagggctcga
      961 attcccgggt cgacatggtc ggcagaagag cactgatcgt actggctcac tcagagagga
     1021 cctccttcaa ctatgccatg aaggaggctg ctgcagcggc tttgaagaag aaaggatggg
     1081 aggtggtgga gtcggacctc tatgccatga acttcaatcc catcatttcc agaaaggaca
     1141 tcacaggtaa actgaaggac cctgcgaact ttcagtatcc tgccgagtct gttctggctt
     1201 ataaagaagg ccatctgagc ccagatattg tggctgaaca aaagaagctg gaagccgcag
     1261 accttgtgat attccagttc cccctgcagt ggtttggagt ccctgccatt ctgaaaggct
     1321 ggtttgagcg agtgttcata ggagagtttg cttacacttt cgctgccatg tatgacaaag
     1381 gacccttccg gagtaagaag gcagtgcttt ccatcaccac tggtggcagt ggctccatgt
     1441 actctctgca agggatccac ggggacatga atgtcattct ctggccaatt cagagtggca
     1501 ttctgcattt ctgtggcttc caagtcttag aacctcaact gacatatagc attgggcaca
     1561 ctccagcaga cgcccgaatt caaatcctgg aaggatggaa gaaacgcctg gagaatattt
     1621 gggatgagac accactgtat tttgctccaa gcagcctctt tgacctaaac ttccaggcag
     1681 gattcttaat gaaaaaagag gtacaggatg aggagaaaaa caagaaattt ggcctttctg
     1741 tgggccatca cttgggcaag tccatcccaa ctgacaacca gatcaaagct agaaaatgag
     1801 cggccgcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     1861 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1921 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     1981 cagtcacgta gcgatagcgg agtgtataat tcttgaagac gaaagggcct cgtgatacgc
     2041 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2101 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2161 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2221 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2281 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2341 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2401 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2461 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2521 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2581 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2641 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2701 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2761 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2821 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2881 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2941 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3001 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3061 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3121 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3361 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3661 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3901 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3961 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4021 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc
     4081 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     4141 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     4201 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     4261 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
     4321 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     4381 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     4441 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     4501 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     4561 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     4621 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
     4681 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
     4741 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
     4801 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
     4861 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
     4921 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
     4981 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
     5041 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     5101 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     5161 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     5221 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     5281 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     5341 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
     5401 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5461 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5521 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
     5581 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
     5641 tgcacggtta cgatgcgccc atctacacca acgtaaccta tcccattacg gtcaatccgc
     5701 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     5761 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt
//
