LOCUS       sequence-330312-        7283 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 7283)
  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 7283)
  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..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1452..1461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1563..1565,1566..2276)
                     /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(1605..1626)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1866..1885)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2213..2234
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3355..3943
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3408..3428)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3826..3837)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3946..3962)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3947..3963)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4030..4506
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4229..4248)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4546..4686
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4546..4675
                     /label=AAV2 ITR
     rep_origin      4761..5216
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4848..4867)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5058..5079
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5233..5252)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5352..5374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5412..5430)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5498..5602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5603..5671,5672..6463)
                     /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(5821..5840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6634..7222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7123..7142
                     /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 gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctcg
     1441 aattcgcggc cgccaccatg ggatgtatca aatcgaaagg taaggacagc ctgtcaggaa
     1501 gtggttgcat taagagcaag ggaaaggatt cgttctgcta tgaaaatgaa gttgccagat
     1561 ctgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1621 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1681 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1741 tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     1801 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1861 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1921 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1981 agctggagta caactggaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     2041 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     2101 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2161 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2221 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagctcg
     2281 agaagatgaa gcggctggtg tacatcagca agatcagcgg ccacctgagc ctggaagaga
     2341 tccagcggat cggcaaggtg tccatcaaga acaaccagcg ggacaacatc accggcgtgc
     2401 tgctgtacct gcagggcctg ttcttccaga tcctggaagg cgagaacgag aaggtggaca
     2461 agctgtacaa gaaaatcctg gtggacgacc ggcacaccaa catcctgtgc ctgaaaaccg
     2521 agtacgacat caccgaccgg atgttcccca actgggccat gaaaaccatc aacctgaacg
     2581 agaacagcga gctgatgatc cagcccatca agagcctgct gcagaccatc acccagagcc
     2641 accgggtgct ggaaaagtac atgcccgcca gagtgatcta cctgatcaac cagggcatca
     2701 accccctgac cgtggagccc cagctggtcg agaagatcat cttcttcagc gacatcctgg
     2761 ccttcagcac cctgaccgag aagctgcccg tgaacgaggt ggtcatcctg gtcaaccggt
     2821 acttcagcat ctgcacccgg atcatcagcg cctacggcgg cgaagtgacc aaatatattg
     2881 gcgattgcgt gatggccagc ttcaccaaag agcagggcga cgccgccatc cggaccagcc
     2941 tggacatcat ctccgagctg aagcagctgc ggcaccacgt ggaggccacc aaccccctgc
     3001 acctgctgta caccggcatc ggcctgagct acggccacgt gatcgagggc aacatgggca
     3061 gcagcctgaa gatggaccac accctgctgg gggacgccgt gaacgtggcc gccaggctgg
     3121 aagccctgac aagacagctg ccctacgccc tggcctttac agccggcgtg aagaagtgct
     3181 gccaggccca gtggaccttc atcaacctgg gagcccacca ggtcaagggc aagcaggaag
     3241 ccatcgaggt ctacaccgtg aatgaggccc agaagtacta cgacaccctg cagatcaccc
     3301 agctgatccg gcagaccctg gaaaacgaca agtagactag ttaagcttat cgataatcaa
     3361 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3421 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3481 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3541 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3601 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3661 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3721 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3781 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3841 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3901 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     3961 cgagacgcga tcgcggcgcg ccattcgatg gcctctctgg cctcgacctc gagtctagag
     4021 atgatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     4081 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     4141 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     4201 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     4261 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     4321 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     4381 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     4441 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     4501 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4561 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4621 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4681 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4741 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4801 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4861 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4921 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4981 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5041 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     5101 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     5161 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     5221 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5281 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5341 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5401 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5461 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5521 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5581 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5641 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5701 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5761 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5821 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5881 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5941 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6001 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6061 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6121 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6181 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6241 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6301 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6361 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6421 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6481 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6541 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6601 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6781 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7021 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7081 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7141 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7201 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7261 ttgctggcct tttgctcaca tgt
//
