LOCUS       SMC1587-pGST2-GS        6694 bp DNA     circular SYN 18-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6694)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 13, 2023 from SnapGene 6.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             918..938
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    945..2678
                     /label=OPTN
     primer_bind     complement(1076..1110)
                     /label=E50K R
     primer_bind     1076..1110
                     /label=E50K F
     CDS             1262..1273
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1455..1495)
                     /label=S173D R
     primer_bind     1455..1495
                     /label=S177D F
     primer_bind     complement(2341..2386)
                     /label=S473D R
     primer_bind     2341..2386
                     /label=S473D F
     misc_feature    2361..2363
                     /label=Feature 17
     promoter        2997..3101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3102..3962
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4133..4721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4965..5042
                     /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."
     CDS             5043..6125
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    6138..6159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6174..6204
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6212..6228
                     /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)."
     primer_bind     6236..6252
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6248..6694
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     complement(6268..6284)
                     /label=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 aatcgatgaa aacctgtatt ttcagggatc aggcatgtcc catcaacctc
      961 tcagctgcct cactgaaaag gaggacagcc ccagtgaaag cacaggaaat ggaccccccc
     1021 acctggccca cccaaacctg gacacgttta ccccggagga gctgctgcag cagatgaaag
     1081 agctcctgac cgagaaccac cagctgaaag aagccatgaa gctaaataat caagccatga
     1141 aagggagatt tgaggagctt tcggcctgga cagagaaaca gaaggaagaa cgccagtttt
     1201 ttgagataca gagcaaagaa gcaaaagagc gtctaatggc cttgagtcat gagaatgaga
     1261 aattgaagga agagcttgga aaactaaaag ggaaatcaga aaggtcatct gaggacccca
     1321 ctgatgactc caggcttccc agggccgaag cggagcagga aaaggaccag ctcaggaccc
     1381 aggtggtgag gctacaagca gagaaggcag acctgttggg catcgtgtct gaactgcagc
     1441 tcaagctgaa ctccagcggc tcctcagaag attcctttgt tgaaattagg atggctgaag
     1501 gagaagcaga agggtcagta aaagaaatca agcatagtcc tgggcccacg agaacagtct
     1561 ccactggcac ggcattgtct aaatatagga gcagatctgc agatggggcc aagaattact
     1621 tcgaacatga ggagttaact gtgagccagc tcctgctgtg cctaagggaa gggaatcaga
     1681 aggtggagag acttgaagtt gcactcaagg aggccaaaga aagagtttca gattttgaaa
     1741 agaaaacaag taatcgttct gagattgaaa cccagacaga ggggagcaca gagaaagaga
     1801 atgatgaaga gaaaggcccg gagactgttg gaagcgaagt ggaagcactg aacctccagg
     1861 tgacatctct gtttaaggag cttcaagagg ctcatacaaa actcagcgaa gctgagctaa
     1921 tgaagaagag acttcaagaa aagtgtcagg cccttgaaag gaaaaattct gcaattccat
     1981 cagagttgaa tgaaaagcaa gagcttgttt atactaacaa aaagttagag ctacaagtgg
     2041 aaagcatgct atcagaaatc aaaatggaac aggctaaaac agaggatgaa aagtccaaat
     2101 taactgtgct acagatgaca cacaacaagc ttcttcaaga acataataat gcattgaaaa
     2161 caattgagga actaacaaga aaagagtcag aaaaagtgga cagggcagtg ctgaaggaac
     2221 tgagtgaaaa actggaactg gcagagaagg ctctggcttc caaacagctg caaatggatg
     2281 aaatgaagca aaccattgcc aagcaggaag aggacctgga aaccatgacc atcctcaggg
     2341 ctcagatgga agtttactgt tctgattttc atgctgaaag agcagcgaga gagaaaattc
     2401 atgaggaaaa ggagcaactg gcattgcagc tggcagttct gctgaaagag aatgatgctt
     2461 tcgaagacgg aggcaggcag tccttgatgg agatgcagag tcgtcatggg gcgagaacaa
     2521 gtgactctga ccagcaggct taccttgttc aaagaggagc tgaggacagg gactggcggc
     2581 aacagcggaa tattccgatt cattcctgcc ccaagtgtgg agaggttctg cctgacatag
     2641 acacgttaca gattcacgtg atggattgca tcatttaact cgagcggccg catcgtgact
     2701 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2761 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2821 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     2881 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2941 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3001 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3061 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3121 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3181 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3241 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3301 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     3361 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3421 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3481 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3541 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3601 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     3661 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3721 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3781 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3841 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3901 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3961 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4021 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4081 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4201 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4261 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4621 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4741 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4801 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4861 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4921 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggt
     4981 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     5041 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     5101 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     5161 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     5221 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     5281 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     5341 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     5401 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     5461 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     5521 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     5581 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     5641 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     5701 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     5761 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     5821 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     5881 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     5941 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     6001 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     6061 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6121 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6181 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6241 aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga
     6301 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     6361 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     6421 atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga
     6481 tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc
     6541 gcccatctac accaacgtaa cctatcccat tacggtcaat ccgccgtttg ttcccacgga
     6601 gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg
     6661 ccagacgcga attatttttg atggcgttgg aatt
//
