pGEX-6p-1-hsG3BP1-mCherry

LOCUS       pGEX-6p-1-hsG3BP1-mCherry           7117 bp DNA     circular SYN 08-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7117)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Apr 17, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7117
                     /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"
     primer_bind     869..891
                     /label=Pgex-5
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             951..971
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             978..2375
                     /codon_start=1
                     /label=hsG3BP
                     /translation="MVMEKPSPLLVGREFVRQYYTLLNQAPDMLHRFYGKNSSYVHGGL
                     DSNGKPADAVYGQKEIHRKVMSQNFTNCHTKIRHVDAHATLNDGVVVQVMGLLSNNNQA
                     LRRFMQTFVLAPEGSVANKFYVHNDIFRYQDEVFGGFVTEPQEESEEEVEEPEERQQTP
                     EVVPDDSGTFYDQAVVSNDMEEHLEEPVAEPEPDPEPEPEQEPVSEIQEEKPEPVLEET
                     APEDAQKSSSPAPADIAQTVQEDLRTFSWASVTSKNLPPSGAVPVTGIPPHVVKVPASQ
                     PRPESKPESQIPPQRPQRDQRVREQRINIPPQRGPRPIREAGEQGDIEPRRMVRHPDSH
                     QLFIGNLPHEVDKSELKDFFQSYGNVVELRINSGGKLPNFGFVVFDDSEPVQKVLSNRP
                     IMFRGEVRLNVEEKKTRAAREGDRRDNRLRGPGGPRGGLGGGMRGPPRGGMVQKPGFGV
                     GRGLAPRQ"
     misc_feature    complement(2358..2396)
                     /label=Primer R in pGEX-6P-1-G3BP1
     misc_feature    2375..2413
                     /label=Primer F for mCherry in N ter-G3BP1
     CDS             2394..3101
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNL
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    complement(3081..3119)
                     /label=Primer R for mCherry in N ter of G3BP1
     misc_feature    3099..3137
                     /label=Primer F in pGEX-6P-1-G3BP1
     primer_bind     complement(3167..3189)
                     /label=pgex-3
     promoter        3420..3524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3525..4385
                     /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      4556..5144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5388..5465
                     /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             5466..6548
                     /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    6561..6582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6597..6627
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6635..6651
                     /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     6659..6675
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6671..7117
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     complement(6691..6707)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcttatcga 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 gaaaacctgt
      961 atttccaggg cggtagtatg gttatggaaa aaccgagtcc gctgctggtt ggtcgcgaat
     1021 ttgtgcgtca gtattatacc ctgctgaatc aggccccgga tatgctgcat cgtttttatg
     1081 gtaaaaatag cagttacgtg catggtggtc tggatagcaa tggtaaaccg gcagatgccg
     1141 tgtatggtca gaaagaaatt catcgtaaag tgatgagtca gaattttacc aattgccata
     1201 ccaaaattcg tcatgttgat gcccatgcaa ccctgaatga tggcgtggtg gtgcaggtta
     1261 tgggtctgct gagtaataat aatcaggcac tgcgccgctt tatgcagacc tttgttctgg
     1321 caccggaagg tagtgttgca aataagtttt atgtgcataa cgatatcttc cgttatcagg
     1381 atgaagtgtt tggcggcttt gtgaccgaac cgcaggaaga aagtgaagaa gaagttgaag
     1441 aaccggaaga acgtcagcag accccggaag ttgtgccgga tgatagtggt accttttatg
     1501 atcaggccgt ggttagcaat gatatggaag aacatctgga agaaccggtt gcagaaccgg
     1561 aaccggaccc tgaaccggaa cctgaacagg aaccggtgag cgaaattcag gaagaaaaac
     1621 cggaaccggt tctggaagaa accgccccgg aagatgccca gaaaagcagt agtccggcac
     1681 cggcagatat tgcccagacc gttcaggaag atctgcgcac ctttagttgg gcaagcgtta
     1741 ccagcaaaaa tctgccgccg agcggtgccg ttccggtgac aggtattccg ccgcatgtgg
     1801 ttaaagttcc ggccagtcag ccgcgtccgg aaagtaaacc ggaaagccag attccgccgc
     1861 agcgtccgca gcgtgatcag cgtgttcgtg aacagcgtat taatattccg ccgcaacgcg
     1921 gcccgcgccc gattcgtgaa gccggtgaac agggcgatat tgaaccgcgt cgtatggtgc
     1981 gtcatccgga tagccatcag ctgtttattg gtaatctgcc gcatgaagtt gataaaagcg
     2041 aactgaaaga tttctttcag agctatggta atgtggtgga actgcgcatt aatagtggtg
     2101 gcaaactgcc gaattttggt tttgtggtgt ttgatgatag tgaaccggtt cagaaagttc
     2161 tgagtaatcg cccgattatg tttcgtggcg aagttcgtct gaatgtggaa gaaaagaaaa
     2221 cccgtgccgc acgtgaaggc gatcgccgcg ataatcgtct gcgtggcccg ggtggcccgc
     2281 gtggtggttt aggtggtggc atgcgcggtc cgccgcgtgg tggcatggtt cagaaaccgg
     2341 gttttggtgt tggtcgcggc ctggccccgc gccagggtgg tagtggtggt agtatggtga
     2401 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     2461 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     2521 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     2581 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     2641 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     2701 acttggagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     2761 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     2821 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     2881 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     2941 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     3001 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     3061 agggccgcca ctccaccggc ggcatggacg agctgtacaa gctcgagcgg ccgcatcgtg
     3121 actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3181 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3241 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     3301 atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     3361 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     3421 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3481 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3541 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3601 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3661 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3721 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     3781 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3841 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3901 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3961 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4021 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     4081 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4141 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4201 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     4261 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     4321 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     4381 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     4441 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4501 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4561 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4621 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4681 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4741 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4801 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4861 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4921 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4981 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5041 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5101 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5161 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5221 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     5281 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     5341 attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat
     5401 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     5461 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     5521 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     5581 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     5641 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     5701 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     5761 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     5821 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     5881 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     5941 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     6001 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     6061 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     6121 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     6181 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     6241 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     6301 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     6361 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     6421 tgcccgtctc actggtgaaa agaaaaacca cgctagcgcc caatacgcaa accgcctctc
     6481 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6541 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta
     6601 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6661 ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg
     6721 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     6781 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     6841 cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg
     6901 cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     6961 tgcgcccatc tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac
     7021 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     7081 aggccagacg cgaattattt ttgatggcgt tggaatt
//