LOCUS       sequence-306856-        7080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2P-activatable bPAC for AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  AUTHORS   Kinjo T, Watabe T, Kobachi K, Terai K, Matsuda M
  TITLE     Single-Cell Activation of the cAMP-Signaling Pathway in 3D Tissues 
            with FRET-Assisted Two-Photon Activation of bPAC.
  JOURNAL   ACS Chem Biol. 2020 Oct 19. doi: 10.1021/acschembio.0c00333.
  PUBMED    33074647
REFERENCE   2  (bases 1 to 7080)
  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: "ACS Chem 
            Biol. 2020 Oct 19. doi: 10.1021/acschembio.0c00333."
COMMENT     Sequence Label: pAAV_2pabPAC
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        208..511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        512..714
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     665..685
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     711..735
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          863..1241
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             3047..3103
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3104..3814
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    3829..4305
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4028..4047)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4343..4483
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4343..4472
                     /label=AAV2 ITR
     rep_origin      4558..5013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4645..4664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4855..4876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5030..5049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5149..5171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5209..5227)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5295..5399
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5400..5468,5469..6260)
                     /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(5618..5637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6431..7019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6920..6939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccaagc gtggagctag ttattaatag taatcaatta cggggtcatt
      181 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      241 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      301 gtcaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      361 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      421 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      481 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      541 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      601 gagtttgttt tgcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      661 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      721 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      781 cgggaccgat ccagcctccg cggattcgaa tcccggccgg gaacggtgca ttggaacgcg
      841 gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc ccacaaaaaa
      901 tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt ccctaatctc
      961 tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt ctaaagaata
     1021 acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat atttctgcat
     1081 ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa tccagctacc
     1141 attctgcttt tattttatgg ttgggataag gctggattat tctgagtcca agctaggccc
     1201 ttttgctaat catgttcata cctcttatct tcctcccaca gctcctgggc aacgtgctgg
     1261 tctgtgtgct ggcccatcac tttggcaaag aattgggatt cgaacatcga ttgaattcac
     1321 catggtgagc aagggcgagg agaccacaat gggcgtaatc aagcccgaca tgaagatcaa
     1381 gctgaagatg gagggcaacg tgaatggcca cgccttcgtg atcgagggcg agggcgaggg
     1441 caagccctac gacggcacca acaccatcaa cctggaggtg aaggagggag cccccctgcc
     1501 cttctcctac gacattctga ccaccgcgtt cgcctacggc aacagggcct tcaccaagta
     1561 ccccgacgac atccccaact acttcaagca gtccttcccc gagggctact cttgggagcg
     1621 caccatgacc ttcgaggaca agggcatcgt gaaggtgaag tccgacatct ccatggagga
     1681 ggactccttc atctacgaga tacacctcaa gggcgagaac ttccccccca acggccccgt
     1741 gatgcagaag aagaccaccg gctgggacgc ctccaccgag aggatgtacg tgcgcgacgg
     1801 cgtgctgaag ggcgacgtca agcacaagct gctgctggag ggcggcggcc accaccgcgt
     1861 tgacttcaag accatctaca gggccaagaa ggcggtgaag ctgcccgact atcactttgt
     1921 ggaccaccgc atcgagatcc tgaaccacga caaggactac aacaaggtga ccgtttacga
     1981 gagcgccgtg gcccgcatga tgaagcggct ggtgtacatc agcaagatca gcggccacct
     2041 gagcctggaa gagatccagc ggatcggcaa ggtgtccatc aagaacaacc agcgggacaa
     2101 catcaccggc gtgctgctgt acctgcaggg cctgttcttc cagatcctgg aaggcgagaa
     2161 cgagaaggtg gacaagctgt acaagaaaat cctggtggac gaccggcaca ccaacatcct
     2221 gtgcctgaaa accgagtacg acatcaccga ccggatgttc cccaactggg ccatgaaaac
     2281 catcaacctg aacgagaaca gcgagctgat gatccagccc atcaagagcc tgctgcagac
     2341 catcacccag agccaccggg tgctggaaaa gtacatgccc gccagagtga tctacctgat
     2401 caaccagggc atcaaccccc tgaccgtgga gccccagctg gtcgagaaga tcatcttctt
     2461 cagcgacatc ctggccttca gcaccctgac cgagaagctg cccgtgaacg aggtggtcat
     2521 cctggtcaac cggtacttca gcatctgcac ccggatcatc agcgcctacg gcggcgaagt
     2581 gaccaagttc atcggcgact gcgtgatggc cagcttcacc aaagagcagg gcgacgccgc
     2641 catccggacc agcctggaca tcatctccga gctgaagcag ctgcggcacc acgtggaggc
     2701 caccaacccc ctgcacctgc tgtacaccgg catcggcctg agctacggcc acgtgatcga
     2761 gggcaacatg ggcagcagcc tgaagatgga ccacaccctg ctgggggacg ccgtgaacgt
     2821 ggccgccagg ctggaagccc tgacaagaca gctgccctac gccctggcct ttacagccgg
     2881 cgtgaagaag tgctgccagg cccagtggac cttcatcaac ctgggagccc accaggtcaa
     2941 gggcaagcag gaagccatcg aggtctacac cgtgaatgag gcccagaagt actacgacac
     3001 cctgcagatc acccagctga tccggcagac cctggaaaac gacaaggcga cgaattttag
     3061 tctactgaaa caagcgggag acgtggagga aaaccctgga cctatggtga gcaagggcga
     3121 ggagaccaca atgggcgtaa tcaagcccga catgaagatc aagctgaaga tggagggcaa
     3181 cgtgaatggc cacgccttcg tgatcgaggg cgagggcgag ggcaagccct acgacggcac
     3241 caacaccatc aacctggagg tgaaggaggg agcccccctg cccttctcct acgacattct
     3301 gaccaccgcg ttcgcctacg gcaacagggc cttcaccaag taccccgacg acatccccaa
     3361 ctacttcaag cagtccttcc ccgagggcta ctcttgggag cgcaccatga ccttcgagga
     3421 caagggcatc gtgaaggtga agtccgacat ctccatggag gaggactcct tcatctacga
     3481 gatacacctc aagggcgaga acttcccccc caacggcccc gtgatgcaga agaagaccac
     3541 cggctgggac gcctccaccg agaggatgta cgtgcgcgac ggcgtgctga agggcgacgt
     3601 caagcacaag ctgctgctgg agggcggcgg ccaccaccgc gttgacttca agaccatcta
     3661 cagggccaag aaggcggtga agctgcccga ctatcacttt gtggaccacc gcatcgagat
     3721 cctgaaccac gacaaggact acaacaaggt gaccgtttac gagagcgccg tggcccgcaa
     3781 ctccaccgac ggcatggacg agctgtacaa gtaactcgag agatctacgg gtggcatccc
     3841 tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc
     3901 cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt
     3961 atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc
     4021 tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc
     4081 tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag
     4141 gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata
     4201 ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa
     4261 ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta
     4321 accacgtgcg gaccgaggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4381 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     4441 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     4501 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     4561 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4621 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4681 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4741 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     4801 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4861 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     4921 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4981 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     5041 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     5101 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     5161 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     5221 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     5281 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5341 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5401 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5461 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5521 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5581 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5641 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5701 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5761 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5821 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5881 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5941 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     6001 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6061 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6121 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6181 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6241 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     6301 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6361 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6421 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6481 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6541 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     6601 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6661 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6721 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6781 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6841 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6901 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6961 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7021 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
