LOCUS       sequence-212384-        6988 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 6988)
  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 6988)
  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..6988
                     /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(2671..2948)
                     /label=chicken beta-actin promoter
     repeat_region   3259..3388
                     /label=AAV2 ITR
     rep_origin      3460..3915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3547..3566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3757..3778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3941..4045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4046..4114,4115..4906)
                     /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(4264..4283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5077..5665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5566..5585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6195..6316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6232..6251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6286..6305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    6359..6947
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6465..6476
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6874..6894
                     /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 cgccctggtg 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 aggcagcgcg cagcgactcc cgcccgccgc gcgcttcgct ttttataggg
     2701 ccgccgccgc cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc tctgattggc
     2761 tgccgccgca cctctccgcc ccgccccccc cccccccccg cccccccccg ccccgcctgg
     2821 cgcgcgcccc cccccccccc ccgcccccat cgctgcacaa aataattaaa aaataaataa
     2881 atacaaaatt gggggtgggg agggggggga gatggggaga gtgaagcaga acgtggggct
     2941 cacctcgacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     3001 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     3061 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3121 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     3181 cattattgac gtcaatggta ctcttcctag agcatggcta cgtagataag tagcatggcg
     3241 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3301 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3361 cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc
     3421 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     3481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3901 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3961 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4021 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4081 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4141 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4201 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4261 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4321 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4381 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4441 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4501 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4561 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4621 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4681 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4741 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4801 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4861 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4921 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4981 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5041 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5101 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5161 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5221 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5281 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5341 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5401 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5461 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5521 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5581 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5641 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5701 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5761 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5821 cgagtcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     5881 cttgtagtta atgattaacc cgccatgcta cccgtattac cgcctttgag tgagctgata
     5941 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgagcga gcgcgcagag
     6001 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
     6061 acttatctac gtagccatgc tctaggaaga tccttatcga ttttaccaca tttgtagagg
     6121 ttttacttgc tttaaaaaac ctcccacatc tccccctgaa cctgaaacat aaaatgaatg
     6181 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6241 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6301 tcatcaatgt atcttatcat gtctgctcga agcggccgcc cgggtcgacg gtatcgatgc
     6361 ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag acgcggaaga
     6421 ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg
     6481 acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag cagccaagga
     6541 aaggacgatg atttccccga caacaccacg gaattgtcag tgcccaacag ccgagcccct
     6601 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg gagggggaaa
     6661 gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg
     6721 tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa ctcctcataa
     6781 agagacagca accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
     6841 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
     6901 agaataccag tcaatctttc acaaattttg taatccagag gttgattatc gataagcttg
     6961 atatcgaatt cgaacctctt cgagggac
//
