LOCUS       sequence-198001-        6989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent, Non-phosphorylatable PKA consensus substrate tagged 
            with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6989)
  AUTHORS   Chen Y, Granger AJ, Tran T, Saulnier JL, Kirkwood A, Sabatini BL
  TITLE     Endogenous Galphaq-Coupled Neuromodulator Receptors Activate Protein
            Kinase A.
  JOURNAL   Neuron. 2017 Dec 6;96(5):1070-1083.e5. doi: 
            10.1016/j.neuron.2017.10.023. Epub 2017 Nov 16.
  PUBMED    29154125
REFERENCE   2  (bases 1 to 6989)
  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"; doi: 
            "10.1016/j.neuron.2017.10.023"; journalName: "Neuron"; date: 
            "2017-12-6- 6"; volume: "96"; issue: "5"; pages: "1070-1083.e5"
COMMENT     Sequence Label: AAV-FLEX-mCherry-FHA1-subT391A-mCherry
FEATURES             Location/Qualifiers
     source          1..6989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..34
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    105..138
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      159..168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             165..872
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1401..2111
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     protein_bind    complement(2125..2158)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(2221..2254)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(2289..2308)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2370..2392)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2672..2949)
                     /label=chicken beta-actin promoter
     repeat_region   3262..3391
                     /label=AAV2 ITR
     rep_origin      3463..3918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3760..3781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3944..4048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4049..4117,4118..4909)
                     /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(4267..4286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5080..5668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5569..5588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5905..6034
                     /label=AAV2 ITR
     polyA_signal    6196..6317
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6233..6252)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6287..6306
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    6360..6948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6466..6477
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6875..6895
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
       61 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
      121 tatcctatac gaagttatac tagaattcgg tacccgccgc caccatggtg agcaagggcg
      181 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      241 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      301 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
      361 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
      421 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      481 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
      541 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
      601 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
      661 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
      721 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
      781 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
      841 actccaccgg cggcatggac gagctgtaca aggctcgcat gcataagttt tcccaggagc
      901 agatcggtga aaatatcgtg tgccgggtca tctgtacaac tggacagatc cccattcgcg
      961 acctgtctgc cgacatcagt caggtgctga aggagaaaag gtctattaag aaagtctgga
     1021 cattcggaag aaaccccgct tgcgactatc acctggggaa cattagcagg ctgtccaata
     1081 agcactttca gattctcctg ggcgaagacg ggaacctcct cctcaacgac atcagcacta
     1141 acgggacctg gttaaatggg caaaaagtcg agaagaacag caatcagtta ctgtctcaag
     1201 gtgatgaaat aaccgttggt gtaggcgtgg aatcagatat tttatctctg gtcattttca
     1261 taaacgacaa atttaagcag tgcctcgagc agaacaaagt tgatcgctct gcaggtaagc
     1321 caggcagcgg cgagggcagc accaagggcc tgcgtcgcgc caccctggtg gacggcggca
     1381 ccggcggctc agaactcatg atggtgagca agggcgagga ggataacatg gccatcatca
     1441 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     1501 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1561 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     1621 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     1681 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     1741 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     1801 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     1861 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     1921 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     1981 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     2041 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2101 tgtacaagta aggggatccc tagtataact tcgtatagca tacattatac gaagttatgg
     2161 gtcgatggtg atgcttggca attcgggtcg atggtgaagc attggtcttc ctattttgat
     2221 ataacttcgt ataggatact ttatacgaag ttatggggat ccaattcttt gccaaaatga
     2281 tgagacagca caataaccag cacgttgccc aggagctgta ggaaaaagaa gaaggcatga
     2341 acatggttag cagaggctct agagccgccg gtcacacgcc agaagccgaa ccccgccctg
     2401 ccccgtcccc cccgaaggca gccgtccccc cgcggacagc cccgagccgc tccccccgca
     2461 caaagggccc tcccggagcc cctcaaggct ttcacgcagc cacagaaaag aaacaagccg
     2521 tcattaaacc aagcgctaat tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg
     2581 tgggagtaac gcggtcagtc agagccgggg cgggcggcgc gaggcggcgg cggagcgggg
     2641 cacggggcga aggcagcgtc gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg
     2701 gccgccgccg ccgccgcctc gccataaaag gaaactttcg gagcgcgccg ctctgattgg
     2761 ctgccgccgc acctctccgc ctcgccccgc cccgcccctc gccccgcccc gccccgcctg
     2821 gcgcgcgccc cccccccccc cccgccccca tcgctgcaca aaataattaa aaaataaata
     2881 aatacaaaat tgggggtggg gagggggggg agatggggag agtgaagcag aacgtggggc
     2941 tcacctcgac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     3001 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     3061 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     3121 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     3181 tcattattga cgtcaatggt acgatcttcc tagagcatgg ctacgtagat aagtagcatg
     3241 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     3301 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3361 gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa tagcgaagag gcccgcaccg
     3421 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     3481 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3901 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3961 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4021 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4081 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4141 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4201 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4261 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4321 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4381 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4441 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4501 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4561 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4621 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4681 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4741 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4801 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4861 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4921 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4981 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5041 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5101 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5161 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5221 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5281 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5341 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5401 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5461 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5521 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5581 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5641 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5701 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5761 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5821 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     5881 gcgttggccg attcattaat gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     5941 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     6001 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
     6061 tacttatcta cgtagccatg ctctaggaag atccttatcg attttaccac atttgtagag
     6121 gttttacttg ctttaaaaaa cctcccacat ctccccctga acctgaaaca taaaatgaat
     6181 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6241 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6301 ctcatcaatg tatcttatca tgtctgctcg aagcggccgc ccgggtcgac ggtatcgatg
     6361 cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag
     6421 aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg
     6481 gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccaagg
     6541 aaaggacgat gatttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc
     6601 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa
     6661 agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc
     6721 gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata
     6781 aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat
     6841 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt
     6901 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat cgataagctt
     6961 gatatcgaat tcgaacctct tcgagggac
//
