LOCUS       sequence-327664-        6038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST fusion vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6038)
  AUTHORS   Tanaka M, Gupta R, Mayer BJ
  TITLE     Differential inhibition of signaling pathways by dominant-negative 
            SH2/SH3 adapter proteins.
  JOURNAL   Mol Cell Biol. 1995 Dec . 15(12):6829-37.
  PUBMED    8524249
REFERENCE   2  (bases 1 to 6038)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol. 1995 Dec . 15(12):6829-37."
COMMENT     Sequence Label: pEBG
FEATURES             Location/Qualifiers
     source          1..6038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    112..133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(148..153,154..171,172..178)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    186..202
                     /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     191..213
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     210..226
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     210..226
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        258..468
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        280..476
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      327..462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        563..1741
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          794..1732
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1689..1709
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1793..2446
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2404..2426
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2453..2470
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3172..3189)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3172..3188)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3181..3203)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3401..3856
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3488..3507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3698..3719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3873..3892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3992..4014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4052..4070)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4138..4242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4243..4311,4312..5103)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4461..4480)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5274..5862
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5763..5782
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6016..6033
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
       61 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      121 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      181 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      241 gcttaattcc ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      301 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      361 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
      421 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
      481 tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
      541 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
      601 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      661 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      721 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      781 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      841 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
      901 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
      961 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
     1021 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     1081 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     1141 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     1201 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     1261 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     1321 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     1381 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     1441 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     1501 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     1561 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     1621 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     1681 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     1741 aggaattctc tagagatccc tcgacctcga gatccattgt gctggatcta caatggcccc
     1801 tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata
     1861 tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa
     1921 caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt
     1981 taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg
     2041 tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag
     2101 atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct
     2161 tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt
     2221 aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt
     2281 atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat
     2341 tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt
     2401 gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggttcc
     2461 gcgtggatcc actagtggta ccatcgatgc ggccgcgact ctagagtgag ggtccccacc
     2521 tgggaccctt gagagtatca ggtctcccac gtgggagaca agaaatccct gtttaatatt
     2581 taaacagcag tgttccccat ctgggtcctt gcacccctca ctctggcctc agccgactgc
     2641 acagcggccc ctgcatcccc ttggctgtga ggcccctgga caagcagagg tggccagagc
     2701 tgggaggcat ggccctgggg tcccacgaat ttgctgggga atctcgtttt tcttcttaag
     2761 acttttggga catggtttga ctcccgaaca tcaccgacgt gtctcctgtt tttctgggtg
     2821 gcctcgggac acctgccctg cccccacgag ggtcaggact gtgactcttt ttagggccag
     2881 gcaggtgcct ggacatttgc cttgctggac ggggactggg gatgtgggag ggagcagaca
     2941 ggaggaatca tgtcaggcct gtgtgtgaaa ggaagctcca ctgtcaccct ccacctcttc
     3001 accccccact caccagtgtc ccctccactg tcacattgta actgaacttc aggataataa
     3061 agtgwtwgcc wccaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3121 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaccgaatt cactggccgt
     3181 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3241 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3301 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     3361 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     3421 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     3481 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3541 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3601 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3661 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3721 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     3781 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3841 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3901 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3961 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4021 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4081 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4141 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4201 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4261 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4321 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4381 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4441 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4501 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4561 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4621 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4681 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4741 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4801 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4861 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4921 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4981 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5041 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5101 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5161 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5221 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5281 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5341 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5401 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5461 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5521 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5581 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5641 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5701 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5761 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5821 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5881 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5941 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     6001 cgaacgaccg agcgcagcga gtcagtgagc gaggaagc
//
