LOCUS       sequence-363589-        7680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of human GST-OSR1-Flag-D164A (kinase-dead).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7680)
  TITLE     Najafov lab plasmids
REFERENCE   2  (bases 1 to 7680)
  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"
COMMENT     Sequence Label: pEBG-hOSR1-Flag-D164A
FEATURES             Location/Qualifiers
     source          1..7680
                     /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"
     CDS             2471..2494
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4103..4126
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4814..4831)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4814..4830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4823..4845)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5043..5498
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5130..5149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5340..5361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5515..5534)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5634..5656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5694..5712)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5780..5884
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5885..5953,5954..6745)
                     /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(6103..6122)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6916..7504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7405..7424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7658..7675
                     /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 gcgtggatct ctggaagttc tgttccaggg gcccctggga tccactagtg gtaccatgtc
     2521 cgaggactcg agcgccctgc cctggtccat caacagggac gattacgagc tgcaggaggt
     2581 gatcgggagt ggagcaactg ctgtagtcca agcagcttat tgtgccccta aaaaggagaa
     2641 agtggcaatc aaacggataa accttgagaa atgtcaaact agcatggatg aactcctgaa
     2701 agaaattcaa gccatgagtc aatgccatca tcctaatatt gtatcttact acacatcttt
     2761 tgtggtaaaa gatgagctgt ggcttgtcat gaagctgcta agtggaggtt ctgttctgga
     2821 tattattaag cacattgtgg caaaagggga acacaaaagt ggagtcctag atgaatctac
     2881 cattgctacg atactccgag aagtactgga agggctggaa tatctgcata aaaatggaca
     2941 gatccacaga gatgtgaaag ctggaaacat tcttcttgga gaagatggct cagtacagat
     3001 tgcagccttt ggggttagtg cttttttagc aactggtggt gatattaccc gaaataaagt
     3061 gagaaagacc tttgttggca ccccttgttg gatggcacct gaagttatgg aacaggtccg
     3121 tggttatgat ttcaaagctg atatttggag ttttggaatt acagcaattg aattggctac
     3181 aggggcggct ccttatcata aatatccacc aatgaaggtt ttaatgctga cactgcagaa
     3241 cgatcctcct tctttggaaa ctggtgttca agataaagaa atgctgaaaa aatatggaaa
     3301 atcatttaga aaaatgattt cattgtgcct tcaaaaagat ccagaaaaaa gaccaacagc
     3361 agcagaacta ttaaggcaca aatttttcca gaaagcaaag aataaagaat ttcttcaaga
     3421 aaaaacattg cagagagcac caaccatttc tgaaagagca aaaaaggttc ggagagtacc
     3481 aggttccagt gggcgtcttc ataagacaga ggatggaggc tgggagtgga gtgatgatga
     3541 atttgatgaa gaaagtgagg aagggaaagc agcaatttca caactcaggt ctccccgagt
     3601 gaaagaatca atatcaaatt ctgagctctt tccaacaact gatcctgtgg gtactttgct
     3661 ccaagttcca gaacagatct ctgctcatct acctcagcca gctgggcaga ttgctacaca
     3721 gccaactcaa gtctctctcc cacccaccgc agagccagca aaaacagctc aggctttgtc
     3781 ttcaggatca ggttcacaag aaaccaagat cccaatcagt ctagtactaa gattaaggaa
     3841 ttccaaaaaa gaactaaatg atattcgatt tgaatttact cctgggagag atacagcaga
     3901 gggtgtctct caggaactca tttctgctgg cctggtcgac ggaagggatt tagtaatagt
     3961 ggcagctaat ttgcagaaaa ttgtggaaga acctcagtca aatcgatctg tcactttcaa
     4021 actggcatct ggtgtcgaag gctcagatat tcctgatgat ggtaaactga taggatttgc
     4081 ccagctcagc atcagcggag gagattacaa ggatgacgat gacaagtagg cggccgcgac
     4141 tctagagtga gggtccccac ctgggaccct tgagagtatc aggtctccca cgtgggagac
     4201 aagaaatccc tgtttaatat ttaaacagca gtgttcccca tctgggtcct tgcacccctc
     4261 actctggcct cagccgactg cacagcggcc cctgcatccc cttggctgtg aggcccctgg
     4321 acaagcagag gtggccagag ctgggaggca tggccctggg gtcccacgaa tttgctgggg
     4381 aatctcgttt ttcttcttaa gacttttggg acatggtttg actcccgaac atcaccgacg
     4441 tgtctcctgt ttttctgggt ggcctcggga cacctgccct gcccccacga gggtcaggac
     4501 tgtgactctt tttagggcca ggcaggtgcc tggacatttg ccttgctgga cggggactgg
     4561 ggatgtggga gggagcagac aggaggaatc atgtcaggcc tgtgtgtgaa aggaagctcc
     4621 actgtcaccc tccacctctt caccccccac tcaccagtgt cccctccact gtcacattgt
     4681 aactgaactt caggataata aagtgtttgc ctccaaaaaa aaaaaaaaaa aaaaaaaaaa
     4741 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4801 aaaaaccgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4861 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4921 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     4981 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     5041 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5101 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5161 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5221 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     5281 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5341 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     5401 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     5461 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     5521 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5581 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5641 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5701 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5761 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5821 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5881 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5941 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6001 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6061 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6121 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6181 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6241 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6301 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6361 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6421 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6481 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6541 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6601 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6661 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6721 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6781 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6841 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7021 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7561 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7621 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
//
