LOCUS       sequence-363831-        7150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of human GST-PKAalpha-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7150)
  TITLE     Najafov lab plasmids
REFERENCE   2  (bases 1 to 7150)
  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-hPKAalpha-Flag
FEATURES             Location/Qualifiers
     source          1..7150
                     /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"
     primer_bind     complement(4284..4301)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4284..4300)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4293..4315)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4513..4968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4600..4619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4810..4831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4985..5004)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5104..5126
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5164..5182)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5250..5354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5355..5423,5424..6215)
                     /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(5573..5592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6386..6974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6875..6894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7128..7145
                     /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 gtaccatggg
     2521 caacgccgcc gccgccaaga agggcagcga gcaggagagc gtgaaagaat tcttagccaa
     2581 agccaaagaa gattttctta aaaaatggga aagtcccgct cagaacacag cccacttgga
     2641 tcagtttgaa cgaatcaaga ccctcggcac gggctccttc gggcgggtga tgctggtgaa
     2701 acacaaggag accgggaacc actatgccat gaagatcctc gacaaacaga aggtggtgaa
     2761 actgaaacag atcgaacaca ccctgaatga aaagcgcatc ctgcaagctg tcaactttcc
     2821 gttcctcgtc aaactcgagt tctccttcaa ggacaactca aacttataca tggtcatgga
     2881 gtacgtgccc ggcggggaga tgttctcaca cctacggcgg atcggaaggt tcagtgagcc
     2941 ccatgcccgt ttctacgcgg cccagatcgt cctgaccttt gagtatctgc actcgctgga
     3001 tctcatctac agggacctga agccggagaa tctgctcatt gaccagcagg gctacattca
     3061 ggtgacagac ttcggtttcg ccaagcgcgt gaagggccgc acttggacct tgtgcggcac
     3121 ccctgagtac ctggcccctg agattatcct gagcaaaggc tacaacaagg ccgtggactg
     3181 gtgggccctg ggggttctta tctatgaaat ggccgctggc tacccgccct tcttcgcaga
     3241 ccagcccatc cagatctatg agaagatcgt ctctgggaag gtgcgcttcc cttcccactt
     3301 cagctctgac ttgaaggacc tgctgcggaa cctcctgcag gtagatctca ccaagcgctt
     3361 tgggaacctc aagaatgggg tcaacgatat caagaaccac aagtggtttg ccacaactga
     3421 ctggattgcc atctaccaga ggaaggtgga agctcccttc ataccaaagt ttaaaggccc
     3481 tggggatacg agtaactttg acgactatga ggaagaagaa atccgggtct ccatcaatga
     3541 gaagtgtggc aaggagtttt ctgagtttgg aggagattac aaggatgacg atgacaatag
     3601 gcggccgcga ctctagagtg agggtcccca cctgggaccc ttgagagtat caggtctccc
     3661 acgtgggaga caagaaatcc ctgtttaata tttaaacagc agtgttcccc atctgggtcc
     3721 ttgcacccct cactctggcc tcagccgact gcacagcggc ccctgcatcc ccttggctgt
     3781 gaggcccctg gacaagcaga ggtggccaga gctgggaggc atggccctgg ggtcccacga
     3841 atttgctggg gaatctcgtt tttcttctta agacttttgg gacatggttt gactcccgaa
     3901 catcaccgac gtgtctcctg tttttctggg tggcctcggg acacctgccc tgcccccacg
     3961 agggtcagga ctgtgactct ttttagggcc aggcaggtgc ctggacattt gccttgctgg
     4021 acggggactg gggatgtggg agggagcaga caggaggaat catgtcaggc ctgtgtgtga
     4081 aaggaagctc cactgtcacc ctccacctct tcacccccca ctcaccagtg tcccctccac
     4141 tgtcacattg taactgaact tcaggataat aaagtgtttg cctccaaaaa aaaaaaaaaa
     4201 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4261 aaaaaaaaaa aaaaaccgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4321 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4381 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4441 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     4501 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4561 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     4621 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4681 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4741 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4801 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     4861 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     4921 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4981 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5041 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5101 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5161 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5221 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5281 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5341 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5401 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5461 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5521 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5581 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5641 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5701 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5761 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5821 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5881 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5941 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6001 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6061 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6121 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6181 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6241 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6301 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6361 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6421 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6481 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6541 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6601 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6661 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6721 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6781 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6841 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6901 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6961 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7021 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7081 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7141 gcgaggaagc
//
