LOCUS       sequence-363613-        6167 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of human PKAalpha-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6167)
  TITLE     Najafov lab plasmids
REFERENCE   2  (bases 1 to 6167)
  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: pcDNA5-FRT-TO-hPKAalpha-Flag
FEATURES             Location/Qualifiers
     source          1..6167
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(2119..2136)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2125..2349
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2482..2501)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2633..2680
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2688..3710
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2688..3710
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    3840..3961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3877..3896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3931..3950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4010..4026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4010..4026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4023..4045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4034..4050
                     /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)."
     promoter        complement(join(4058..4064,4065..4082,4083..4088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4103..4124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4241..4258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4412..5000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4492..4511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5171..5962,5963..6031))
                     /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     5794..5813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6032..6136)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca tgggcaacgc cgccgccgcc aagaagggca
     1021 gcgagcagga gagcgtgaaa gaattcttag ccaaagccaa agaagatttt cttaaaaaat
     1081 gggaaagtcc cgctcagaac acagcccact tggatcagtt tgaacgaatc aagaccctcg
     1141 gcacgggctc cttcgggcgg gtgatgctgg tgaaacacaa ggagaccggg aaccactatg
     1201 ccatgaagat cctcgacaaa cagaaggtgg tgaaactgaa acagatcgaa cacaccctga
     1261 atgaaaagcg catcctgcaa gctgtcaact ttccgttcct cgtcaaactc gagttctcct
     1321 tcaaggacaa ctcaaactta tacatggtca tggagtacgt gcccggcggg gagatgttct
     1381 cacacctacg gcggatcgga aggttcagtg agccccatgc ccgtttctac gcggcccaga
     1441 tcgtcctgac ctttgagtat ctgcactcgc tggatctcat ctacagggac ctgaagccgg
     1501 agaatctgct cattgaccag cagggctaca ttcaggtgac agacttcggt ttcgccaagc
     1561 gcgtgaaggg ccgcacttgg accttgtgcg gcacccctga gtacctggcc cctgagatta
     1621 tcctgagcaa aggctacaac aaggccgtgg actggtgggc cctgggggtt cttatctatg
     1681 aaatggccgc tggctacccg cccttcttcg cagaccagcc catccagatc tatgagaaga
     1741 tcgtctctgg gaaggtgcgc ttcccttccc acttcagctc tgacttgaag gacctgctgc
     1801 ggaacctcct gcaggtagat ctcaccaagc gctttgggaa cctcaagaat ggggtcaacg
     1861 atatcaagaa ccacaagtgg tttgccacaa ctgactggat tgccatctac cagaggaagg
     1921 tggaagctcc cttcatacca aagtttaaag gccctgggga tacgagtaac tttgacgact
     1981 atgaggaaga agaaatccgg gtctccatca atgagaagtg tggcaaggag ttttctgagt
     2041 ttggaggaga ttacaaggat gacgatgaca ataggcggcc gctcgagtct agagggcccg
     2101 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2161 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2221 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2281 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2341 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2401 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2461 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2521 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2581 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc tagaagttcc
     2641 tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa aagcctgaac
     2701 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     2761 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     2821 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     2881 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     2941 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     3001 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     3061 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     3121 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     3181 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     3241 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     3301 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     3361 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     3421 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     3481 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     3541 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     3601 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     3661 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtactac
     3721 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3781 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3961 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5461 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5581 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
