LOCUS       sequence-225608-        7138 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Alternative spliced variant of the mouse PKA catalytic subunit alpha
            C-terminally tagged by monomeric paGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  AUTHORS   Tillo SE, Xiong WH, Takahashi M, Miao S, Andrade AL, Fortin DA, Yang
            G, Qin M, Smoody BF, Stork PJS, Zhong H
  TITLE     Liberated PKA Catalytic Subunits Associate with the Membrane via 
            Myristoylation to Preferentially Phosphorylate Membrane Substrates.
  JOURNAL   Cell Rep. 2017 Apr 18;19(3):617-629. doi: 
            10.1016/j.celrep.2017.03.070.
  PUBMED    28423323
REFERENCE   2  (bases 1 to 7138)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2017.03"; journalName: "Cell Rep"; date: 
            "2017-04-18- 18"; volume: "19"; issue: "3"; pages: "617-629"
COMMENT     Sequence Label: pCDNA3_altPKA-C-alpha-mPAGFP
FEATURES             Location/Qualifiers
     source          1..7138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        468..671
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     622..642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     716..735
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        716..734
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1785..1811
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             join(1812..1814,1815..2525)
                     /codon_start=1
                     /product="photoactivatable GFP variant (Patterson et al., 
                     2002)"
                     /label=PA-GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1854..1875)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2115..2134)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2462..2483
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2544..2562)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2545..2562)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2582..2599)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2588..2812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2858..3286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2945..2964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3155..3176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3295..3315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3300..3629
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3480..3615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3542..3561
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3696..4490
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3750..3769)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4360..4379
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4664..4785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4701..4720)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4755..4774
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4834..4850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4834..4850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4847..4869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4858..4874
                     /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(4882..4888,4889..4906,4907..4912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4927..4948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5065..5082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5236..5824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5316..5335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5995..6786,6787..6855))
                     /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     6618..6637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6856..6960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7035..7054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
       61 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
      121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      661 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      721 cgactcacta tagggagacc caagctacca tggcttccag ctccaacgat gtgaaagagt
      781 tcctagccaa agccaaggaa gatttcctga aaaaatggga gaccccttct cagaatacag
      841 cccagttgga tcagtttgat agaatcaaga cccttggcac cggctccttt gggcgagtga
      901 tgctggtgaa gcacaaggag agtgggaacc actacgccat gaagatctta gacaagcaga
      961 aggtggtgaa gctaaagcag atcgagcaca ctctgaatga gaagcgcatc ctgcaggccg
     1021 tcaacttccc gttcctggtc aaacttgaat tctccttcaa ggacaactca aacctgtaca
     1081 tggtcatgga gtatgtagct ggtggcgaga tgttctccca cctacggcgg attggaaggt
     1141 tcagcgagcc ccatgcccgt ttctacgcgg cgcagatcgt cctgaccttt gagtatctgc
     1201 actccctgga cctcatctac cgggacctga agcccgagaa tcttctcatc gaccagcagg
     1261 gctatattca ggtgacagac ttcggttttg ccaagcgtgt gaaaggccgt acttggacct
     1321 tgtgtgggac ccctgagtac ttggcccccg agattatcct gagcaaaggc tacaacaagg
     1381 ctgtggactg gtgggctctc ggagtcctca tctacgagat ggctgctggt tacccaccct
     1441 tcttcgctga ccagcctatc cagatctatg agaaaatcgt ctctgggaag gtgcggttcc
     1501 catcccactt cagctctgac ttgaaggacc tgctgcggaa ccttctgcaa gtggatctca
     1561 ccaagcgctt tgggaacctc aagaacgggg tcaatgacat caagaaccac aagtggtttg
     1621 ccacgactga ctggattgcc atctatcaga gaaaggtgga agctcccttc ataccaaagt
     1681 ttaaaggccc tggggacacg agtaactttg acgactatga ggaggaagag atccgggtct
     1741 ccatcaatga gaagtgtggc aaggagttta ctgagtttat cgattatgat gttcctgatt
     1801 atgcaagctt aatgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1861 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1921 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1981 ggcccaccct cgtgaccacc ttcagctacg gcgtgcagtg cttcagccgc taccccgacc
     2041 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2101 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2161 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2221 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2281 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2341 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2401 acaaccacta cctgagccac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     2461 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2521 acaagtaatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     2581 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2641 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2701 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2761 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2821 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2881 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2941 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3001 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3061 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3121 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3181 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3241 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3301 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3421 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3481 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3541 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3601 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3661 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3721 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3781 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3841 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3901 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3961 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4021 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4081 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4141 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4201 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4261 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4321 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4381 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4441 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4501 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4561 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4621 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4681 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4741 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4801 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4861 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4921 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4981 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5041 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5101 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5161 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5221 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5281 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5341 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5401 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5461 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5521 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5581 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5641 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5701 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5761 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5821 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5881 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5941 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6001 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6061 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6121 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6181 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6241 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6301 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6361 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6421 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6481 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6541 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6601 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6661 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6721 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6781 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6841 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6901 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6961 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7021 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7081 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggc
//
