LOCUS       sequence-321719-        7286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuronal expression of PACmn with dark Venus, a photoactivatable 
            adenylyl cyclase derived from bPAC, membrane-anchored, no/reduced 
            dark activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7286)
  AUTHORS   Yang S, Constantin OM, Sachidanandan D, Hofmann H, Kunz TC, 
            Kozjak-Pavlovic V, Oertner TG, Nagel G, Kittel RJ, Gee CE, Gao S
  TITLE     PACmn for improved optogenetic control of intracellular cAMP.
  JOURNAL   BMC Biol. 2021 Oct 18;19(1):227. doi: 10.1186/s12915-021-01151-9.
  PUBMED    34663304
REFERENCE   2  (bases 1 to 7286)
  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: "BMC Biol.";
            date: "2021-10-18"; volume: "19(1):227. doi"; pages: " 
            10.1186/s12915-021-01151-9"
COMMENT     Sequence Label: pAAV-CaMKII-2xLyn-ERex-Venus(Y145W)-bPAC(F198Y)
FEATURES             Location/Qualifiers
     source          1..7286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..1293
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1308..1317
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1419..1421,1422..2132)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1461..1482)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1722..1741)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2069..2090
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3210..3798
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3263..3283)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3681..3692)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3801..3817)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3802..3818)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3878..4354
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4077..4096)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4394..4534
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4394..4523
                     /label=AAV2 ITR
     rep_origin      4609..5064
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4696..4715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4906..4927
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5081..5100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5200..5222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5260..5278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5346..5450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5451..5519,5520..6311)
                     /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(5669..5688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6482..7070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6971..6990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7132..7272
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7143..7272
                     /label=AAV2 ITR
ORIGIN
        1 ttaacattat ggccttaggt cacttcatct ccatggggtt cttcttctga ttttctagaa
       61 aatgagatgg gggtgcagag agcttcctca gtgacctgcc cagggtcaca tcagaaatgt
      121 cagagctaga acttgaactc agattactaa tcttaaattc catgccttgg gggcatgcaa
      181 gtacgatata cagaaggagt gaactcatta gggcagatga ccaatgagtt taggaaagaa
      241 gagtccaggg cagggtacat ctacaccacc cgcccagccc tgggtgagtc cagccacgtt
      301 cacctcatta tagttgcctc tctccagtcc taccttgacg ggaagcacaa gcagaaactg
      361 ggacaggagc cccaggagac caaatcttca tggtccctct gggaggatgg gtggggagag
      421 ctgtggcaga ggcctcagga ggggccctgc tgctcagtgg tgacagatag gggtgagaaa
      481 gcagacagag tcattccgtc agcattctgg gtctgtttgg tacttcttct cacgctaagg
      541 tggcggtgtg atatgcacaa tggctaaaaa gcagggagag ctggaaagaa acaaggacag
      601 agacagaggc caagtcaacc agaccaattc ccagaggaag caaagaaacc attacagaga
      661 ctacaagggg gaagggaagg agagatgaat tagcttcccc tgtaaacctt agaacccagc
      721 tgttgccagg gcaacggggc aatacctgtc tcttcagagg agatgaagtt gccagggtaa
      781 ctacatcctg tctttctcaa ggaccatccc agaatgtggc acccactagc cgttaccata
      841 gcaactgcct ctttgcccca cttaatccca tcccgtctgt taaaagggcc ctatagttgg
      901 aggtggggga ggtaggaaga gcgatgatca cttgtggact aagtttgttc gcatcccctt
      961 ctccaacccc ctcagtacat caccctgggg gaacagggtc cacttgctcc tgggcccaca
     1021 cagtcctgca gtattgtgta tataaggcca gggcaaagag gagcaggttt taaagtgaaa
     1081 ggcaggcagg tgttggggag gcagttaccg gggcaacggg aacagggcgt ttcggaggtg
     1141 gttgccatgg ggacctggat gctgacgaag gctcgcgagg ctgtgagcag ccacagtgcc
     1201 ctgctcagaa gccccaagct cgtcagtcaa gccggttctc cgtttgcact caggagcacg
     1261 ggcaggcgag tggcccctag ttctgggggc agctcgaatt cgcggccgcc accatgggat
     1321 gtatcaaatc gaaaggtaag gacagcctgt caggaagtgg ttgcattaag agcaagggaa
     1381 aggattcgtt ctgctatgaa aatgaagttg ccagatctgt gagcaagggc gaggagctgt
     1441 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1501 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
     1561 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc
     1621 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1681 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1741 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     1801 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tggaacagcc
     1861 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     1921 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1981 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
     2041 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2101 ggatcactct cggcatggac gagctgtaca agctcgagaa gatgaagcgg ctggtgtaca
     2161 tcagcaagat cagcggccac ctgagcctgg aagagatcca gcggatcggc aaggtgtcca
     2221 tcaagaacaa ccagcgggac aacatcaccg gcgtgctgct gtacctgcag ggcctgttct
     2281 tccagatcct ggaaggcgag aacgagaagg tggacaagct gtacaagaaa atcctggtgg
     2341 acgaccggca caccaacatc ctgtgcctga aaaccgagta cgacatcacc gaccggatgt
     2401 tccccaactg ggccatgaaa accatcaacc tgaacgagaa cagcgagctg atgatccagc
     2461 ccatcaagag cctgctgcag accatcaccc agagccaccg ggtgctggaa aagtacatgc
     2521 ccgccagagt gatctacctg atcaaccagg gcatcaaccc cctgaccgtg gagccccagc
     2581 tggtcgagaa gatcatcttc ttcagcgaca tcctggcctt cagcaccctg accgagaagc
     2641 tgcccgtgaa cgaggtggtc atcctggtca accggtactt cagcatctgc acccggatca
     2701 tcagcgccta cggcggcgaa gtgaccaaat atattggcga ttgcgtgatg gccagcttca
     2761 ccaaagagca gggcgacgcc gccatccgga ccagcctgga catcatctcc gagctgaagc
     2821 agctgcggca ccacgtggag gccaccaacc ccctgcacct gctgtacacc ggcatcggcc
     2881 tgagctacgg ccacgtgatc gagggcaaca tgggcagcag cctgaagatg gaccacaccc
     2941 tgctggggga cgccgtgaac gtggccgcca ggctggaagc cctgacaaga cagctgccct
     3001 acgccctggc ctttacagcc ggcgtgaaga agtgctgcca ggcccagtgg accttcatca
     3061 acctgggagc ccaccaggtc aagggcaagc aggaagccat cgaggtctac accgtgaatg
     3121 aggcccagaa gtactacgac accctgcaga tcacccagct gatccggcag accctggaaa
     3181 acgacaagta aactagtaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     3241 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3301 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     3361 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     3421 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3481 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3541 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3601 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3661 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3721 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3781 tttgggccgc ctccccgcat cgataccgtc gacctcgaga cgcgatcgcg gcgcgccggc
     3841 ctctctggcc tcgacctcga gtctagagat gatctacggg tggcatccct gtgacccctc
     3901 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3961 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     4021 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     4081 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     4141 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     4201 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     4261 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     4321 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     4381 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4441 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4501 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4561 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4621 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4681 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4741 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4801 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4861 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4921 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     4981 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5041 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     5101 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5161 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5221 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5281 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5341 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5401 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5461 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5521 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5581 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5641 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5701 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5761 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5821 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5881 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5941 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6001 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6061 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6121 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6181 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6241 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6301 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6361 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6421 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6481 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6541 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6601 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6661 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6721 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6781 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6841 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6901 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6961 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7021 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7081 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc
     7141 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
     7201 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     7261 ctaggggttc ctgcggccgc acgcgt
//
