pGEX-2T-G3BP1-Flag

LOCUS       pGEX-2T-G3BP1-Flag               6375 bp DNA     circular SYN 23-NOV-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6375)
  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..6375
                     /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..2366
                     /codon_start=1
                     /label=hsG3BP1 no codon optimzed
                     /translation="MVMEKPSPLLVGREFVRQYYTLLNQAPDMLHRFYGKNSSYVHGGL
                     DSNGKPADAVYGQKEIHRKVMSQNFTNCHTKIRHVDAHATLNDGVVVQVMGLLSNNNQA
                     LRRFMQTFVLAPEGSVANKFYVHNDIFRYQDEVFGGFVTEPQEESEEEVEEPEERQQTP
                     EVVPDDSGTFYDQAVVSNDMEEHLEEPVAEPEPDPEPEPEQEPVSEIQEEKPEPVLEET
                     APEDAQKSSSPAPADIAQTVQEDLRTFSWASVTSKNLPPSGAVPVTGIPPHVVKVPASQ
                     PRPESKPESQIPPQRPQRDQRVREQRINIPPQRGPRPIREAGEQGDIEPRRMVRHPDSH
                     QLFIGNLPHEVDKSELKDFFQSYGNVVELRINSGGKLPNFGFVVFDDSEPVQKVLSNRP
                     IMFRGEVRLNVEEKKTRAAREGDRRDNRLRGPGGPRGGLGGGMRGPPRGGMVQKPGFGV
                     GRGLAPRQDYKDDDDK"
     misc_feature    complement(2322..2363)
                     /label=Primer R for hsG3BP1-flag
     CDS             2340..2363
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2343..2384
                     /label=Primer F for hsG3BP1-Flag
     promoter        2678..2782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2783..3643
                     /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      3814..4402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4646..4723
                     /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             4724..5806
                     /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    5819..5840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5855..5885
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5893..5909
                     /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     5917..5933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5929..6375
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     complement(5949..5965)
                     /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 catggtgatg gagaagccta
      961 gtcccctgct ggtcgggcgg gaatttgtga gacagtatta cacactgctg aaccaggccc
     1021 cagacatgct gcatagattt tatggaaaga actcttctta tgtccatggg ggattggatt
     1081 caaatggaaa gccagcagat gcagtctacg gacagaaaga aatccacagg aaagtgatgt
     1141 cacaaaactt caccaactgc cacaccaaga ttcgccatgt tgatgctcat gccacgctaa
     1201 atgatggtgt ggtagtccag gtgatggggc ttctctctaa caacaaccag gctttgagga
     1261 gattcatgca aacgtttgtc cttgctcctg aggggtctgt tgcaaataaa ttctatgttc
     1321 acaatgatat cttcagatac caagatgagg tctttggtgg gtttgtcact gagcctcagg
     1381 aggagtctga agaagaagta gaggaacctg aagaaagaca gcaaacacct gaggtggtac
     1441 ctgatgattc tggaactttc tatgatcagg cagttgtcag taatgacatg gaagaacatt
     1501 tagaggagcc tgttgctgaa ccagagcctg atcctgaacc agaaccagaa caagaacctg
     1561 tatctgaaat ccaagaggaa aagcctgagc cagtattaga agaaactgcc cctgaggatg
     1621 ctcagaagag ttcttctcca gcacctgcag acatagctca gacagtacag gaagacttga
     1681 ggacattttc ttgggcatct gtgaccagta agaatcttcc acccagtgga gctgttccag
     1741 ttactgggat accacctcat gttgttaaag taccagcttc acagccccgt ccagagtcta
     1801 agcctgaatc tcagattcca ccacaaagac ctcagcggga tcaaagagtg cgagaacaac
     1861 gaataaatat tcctccccaa aggggaccca gaccaatccg tgaggctggt gagcaaggtg
     1921 acattgaacc ccgaagaatg gtgagacacc ctgacagtca ccaactcttc attggcaacc
     1981 tgcctcatga agtggacaaa tcagagctta aagatttctt tcaaagttat ggaaacgtgg
     2041 tggagttgcg cattaacagt ggtgggaaat tacccaattt tggttttgtt gtgtttgatg
     2101 attctgagcc tgttcagaaa gtccttagca acaggcccat catgttcaga ggtgaggtcc
     2161 gtctgaatgt cgaagagaag aagactcgag ctgccaggga aggcgaccga cgagataatc
     2221 gccttcgggg acctggaggc cctcgaggtg ggctgggtgg tggaatgaga ggccctcccc
     2281 gtggaggcat ggtgcagaaa ccaggatttg gagtgggaag ggggcttgcg ccacggcagg
     2341 attacaagga cgacgatgac aagtgagaat tcatcgtgac tgactgacga tctgcctcgc
     2401 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2461 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2521 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt ataattcttg
     2581 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2641 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2701 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2761 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2821 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2881 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2941 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3001 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     3061 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3121 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3181 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3241 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3301 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     3361 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3421 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3481 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3541 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3601 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3661 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3721 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3781 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3841 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3901 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3961 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4021 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4081 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4141 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4201 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4261 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4321 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4381 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4441 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4501 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4561 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4621 cggtatttca caccgcataa attccgacac catcgaatgg tgcaaaacct ttcgcggtat
     4681 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     4741 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     4801 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     4861 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     4921 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     4981 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     5041 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     5101 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     5161 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     5221 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     5281 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     5341 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     5401 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     5461 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     5521 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     5581 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     5641 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     5701 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5761 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5821 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5881 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5941 acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     6001 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     6061 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
     6121 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
     6181 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgta
     6241 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
     6301 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
     6361 gatggcgttg gaatt
//