pGEX-2T-BRE5

LOCUS       pGEX-2T-BRE5                6498 bp DNA     circular SYN 11-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Apr 16, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6498
                     /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"
     CDS             942..2489
                     /codon_start=1
                     /label=BRE5 synthesis codon
                     /translation="MGVTVQDICFAFLQNYYERMRTDPSKLAYFYASTAELTHTNYQSK
                     STNEKDDVLPTVKVTGRENINKFFSRNDAKVRSLKLKLDTIDFQYTGHLHKSILIMATG
                     EMFWTGTPVYKFCQTFILLPSSNGSTFDITNDIIRFISNSFKPYVLTDASLSQSNEENS
                     VSAVEEDKIRHESGVEKEKEKEKSPEISKPKAKKETVKDTTAPTESSTQEKPIVDHSQP
                     RAIPVTKESKIHTETVPSSTKGNHKQDEVSTEELGNVTKLNEKSHKAEKKAAPIKTKEG
                     SVEAINAVNNSSLPNGKEVSDEKPVPGGVKEAETEIKPIEPQVSDAKESGNNASTPSSS
                     PEPVANPPKMTWASKLMNENSDRISKNNTTVEYIRPETLPKKPTERKFEMGNRRDNASA
                     NSKNKKKPVFSTVNKDGFYPIYIRGTNGLREEKLRSALEKEFGKVMRITAADNFAVVDF
                     ETQKSQIDALEKKKKSIDGIEVCLERKTVKKPTSNNPPGIFTNGTRSHRKQPLKRKD"
     misc_feature    1795..1812
                     /label=test Primer 1
     primer_bind     complement(2548..2570)
                     /label=Pgex-3
     promoter        2801..2905
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2906..3766
                     /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      3937..4525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4769..4846
                     /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             4847..5929
                     /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    5942..5963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5978..6008
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6016..6032
                     /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     6040..6056
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6052..6498
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     complement(6072..6088)
                     /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 atcggatgaa aacctgtatt ttcagggatc catgggtgtt accgtgcagg
      961 atatttgttt tgcctttctg cagaattatt acgaacgcat gcgtaccgat ccgagcaaac
     1021 tggcatattt ttatgccagt accgccgaac tgacccatac caattatcag agcaaaagta
     1081 ccaatgaaaa ggatgatgtg ctgccgaccg tgaaagttac cggccgcgaa aatattaata
     1141 agtttttcag ccgcaacgat gcaaaagttc gtagtctgaa actgaaactg gataccattg
     1201 attttcagta taccggtcat ctgcataaaa gcattctgat tatggccacc ggtgaaatgt
     1261 tttggaccgg taccccggtg tataaatttt gtcagacctt tattctgctg ccgagcagta
     1321 atggcagtac ctttgatatt accaatgata ttatccgctt cattagcaat agttttaagc
     1381 cgtatgttct gaccgatgcc agtctgagtc agagcaatga agaaaatagc gttagcgccg
     1441 ttgaagaaga taaaattcgt catgaaagtg gcgttgaaaa agaaaaagaa aaggaaaaaa
     1501 gcccggaaat tagcaaaccg aaagcaaaaa aggaaaccgt gaaagatacc accgccccga
     1561 ccgaaagcag cacccaggaa aaaccgattg tggatcatag ccagccgcgt gcaattccgg
     1621 tgaccaaaga aagtaaaatt cataccgaaa ccgttccgag cagcaccaaa ggtaatcata
     1681 aacaggatga agttagcacc gaagaactgg gcaatgttac caaactgaat gaaaaaagcc
     1741 ataaagccga aaagaaagca gccccgatta agaccaaaga aggtagcgtt gaagccatta
     1801 atgcagtgaa taatagtagt ctgccgaatg gcaaagaagt gagtgatgaa aaaccggttc
     1861 cgggtggtgt taaagaagca gaaaccgaaa ttaagccgat tgaaccgcag gtgagcgatg
     1921 caaaagaaag cggcaataat gccagtaccc cgagtagcag cccggaaccg gttgcaaatc
     1981 cgccgaaaat gacctgggca agtaaactga tgaatgaaaa tagcgatcgc attagtaaaa
     2041 acaataccac cgttgaatac attcgcccgg aaaccctgcc gaaaaaaccg accgaacgca
     2101 aatttgaaat gggcaatcgt cgtgataatg ccagtgccaa tagtaaaaat aagaaaaaac
     2161 cggtgttcag taccgtgaat aaggatggct tttatccgat ctatattcgt ggtaccaatg
     2221 gcctgcgcga agaaaaactg cgtagtgcac tggaaaaaga atttggtaaa gtgatgcgca
     2281 ttaccgccgc agataatttt gccgtggtgg attttgaaac ccagaaaagt cagattgatg
     2341 ccctggaaaa gaaaaagaaa agtattgatg gtatcgaggt gtgtctggaa cgtaaaaccg
     2401 tgaaaaaacc gacaagcaat aatccgccgg gtatttttac caatggtacc cgcagccatc
     2461 gtaaacagcc gctgaaacgt aaagattaag aattcatcgt gactgactga cgatctgcct
     2521 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2581 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2641 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtataattc
     2701 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     2761 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2821 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2881 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2941 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3001 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3061 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     3121 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     3181 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     3241 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3301 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3361 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3421 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
     3481 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3541 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3601 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3661 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3721 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3781 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3841 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3901 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3961 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4021 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4081 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4141 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4201 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4261 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4321 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4381 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4441 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4501 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4561 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4621 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4681 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     4741 gtgcggtatt tcacaccgca taaattccga caccatcgaa tggtgcaaaa cctttcgcgg
     4801 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     4861 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     4921 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     4981 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     5041 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     5101 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     5161 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     5221 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     5281 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     5341 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     5401 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     5461 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     5521 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     5581 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     5641 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     5701 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     5761 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     5821 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5881 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5941 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     6001 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     6061 attacggatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     6121 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     6181 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg
     6241 tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat
     6301 actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac
     6361 gtaacctatc ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt
     6421 tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt
     6481 tttgatggcg ttggaatt
//