LOCUS       sequence-332592-        7301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse PKA regulatory subunit II-alpha 
            C-terminally tagged by sREACh..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7301)
  AUTHORS   Xiong WH, Qin M, Zhong H
  TITLE     Myristoylation alone is sufficient for PKA catalytic subunits to 
            associate with the plasma membrane to regulate neuronal functions.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Apr 13;118(15). pii: 2021658118. doi:
            10.1073/pnas.2021658118.
  PUBMED    33876760
REFERENCE   2  (bases 1 to 7301)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-04-13"; volume: "118(15). pii: 
            2021658118. doi"; pages: " 10.1073/pnas.2021658118"
COMMENT     Sequence Label: pCDNA3_PKA-RII-alpha-sREACh
FEATURES             Location/Qualifiers
     source          1..7301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(130..718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(210..229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(889..1680,1681..1749))
                     /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     1512..1531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1750..1854)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1929..1948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2120..2499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2500..2703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2654..2674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2748..2767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2748..2766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4004..4006,4007..4009,4010..4720)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSVNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(4049..4070)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4310..4329)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(4739..4757)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4740..4757)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4777..4794)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4783..5007
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5053..5481
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5140..5159)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5350..5371
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5490..5510)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5495..5824
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5675..5810
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5737..5756
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5891..6685
                     /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(5945..5964)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6555..6574
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6859..6980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6896..6915)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6950..6969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7029..7045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7029..7045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7042..7064)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7053..7069
                     /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(7077..7083,7084..7101,7102..7107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7122..7143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7260..7277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
       61 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      121 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      181 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
      241 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
      301 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
      361 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
      421 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      481 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      541 gtggtggccc aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      601 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      661 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      721 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      781 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      841 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      961 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1081 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1321 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1381 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1561 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1741 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1861 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     1921 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     1981 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     2041 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     2101 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     2161 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2221 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2281 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2341 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2401 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2461 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2521 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2581 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2641 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2701 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     2761 tatagggaga cccaagcttg cggccgccac catgagccac atccagatcc ccgcggggct
     2821 cacggagctg ctgcagggct acaccgtgga ggtgcttcgg cagcagccgc ccgacctcgt
     2881 cgacttcgcg gtggagtact tcacacgcct gcgcgaggcc cgccgccagg aatcagacac
     2941 gttcatcgtc tccccgacga cctttcacac gcaggagtcc agcgcagtcc ccgtcatcga
     3001 ggaggacggg gagagtgact cggactcgga agatgccgat ctggaagttc cggttcctag
     3061 caaatttact agacgagtat cagtctgtgc agaaacgttt aaccctgatg aagaagagga
     3121 ggataacgat ccaagggtgg ttcatcccaa aactgatgag cagagatgcc ggcttcagga
     3181 agcttgtaaa gatattcttc ttttcaaaaa ccttgatcag gaacagcttt ctcaagttct
     3241 ggatgccatg tttgaaaaga ttgtcaaaac tgacgagcat gtcattgacc aaggcgacga
     3301 cggggacaac ttttatgtca tagaacgggg aacctatgac attttagtaa cgaaagataa
     3361 tcaaacacgt tctgttggtc agtatgacaa ccgtggcagt tttggagaac tagctctgat
     3421 gtacaatacc ccgagagctg ctaccatcat cgccacctca gaaggctccc tttggggatt
     3481 ggaccgggtg acttttagaa gaatcatagt gaaaaacaat gcaaagaaga ggaagatgtt
     3541 tgaatcattt attgagtctg ttccactctt taagtcacta gagatgtcag aacgaatgaa
     3601 gattgtggat gtgatcgggg aaaagatcta taaggacgga gagcgaataa tcgcacaggg
     3661 tgaaaaggcc gacagcttct atatcataga gtctggggaa gtgagcatct tgattagaag
     3721 caagaccaag tcaaacaaga atggagggaa ccaggaggtc gagattgccc attgccataa
     3781 ggggcagtac ttcggagaac ttgccctggt caccaacaaa cccagagctg cttccgctta
     3841 cgcggttgga gatgtcaaat gcttagttat ggatgttcaa gcatttgaga ggcttctggg
     3901 cccctgcatg gacatcatga agaggaacat ctcacattac gaagaacagc tggtgaagat
     3961 gtttggctcc aacttggatc tgatggaccc cgggcaggat atcatggtga gcaagggcga
     4021 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4081 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     4141 gctcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg
     4201 ctacggcctg atgtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     4261 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     4321 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     4381 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactg
     4441 gaacagcgtc aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     4501 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     4561 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     4621 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agagagtgac
     4681 cgccgccggg atcactctcg gcatggacga gctgtacaag taatctagag ggccctattc
     4741 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     4801 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4861 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4921 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4981 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     5041 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5101 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5161 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5221 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5281 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5341 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5401 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5461 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5521 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5581 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5641 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5701 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5761 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5821 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     5881 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5941 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6001 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6061 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6121 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     6181 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     6241 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     6301 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6361 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     6421 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6481 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6541 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6601 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6661 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6721 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6781 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6841 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     6901 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6961 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     7021 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7081 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7141 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7261 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc g
//
