LOCUS       sequence-321718-        6470 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 6470)
  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 6470)
  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-Syn-2xLyn-ERex-Venus(Y145W)-bPAC(F198Y)
FEATURES             Location/Qualifiers
     source          1..6470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..474
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      489..498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(600..602,603..1313)
                     /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(642..663)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(903..922)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1250..1271
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2391..2979
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2444..2464)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2862..2873)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2982..2998)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2983..2999)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3059..3535
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3258..3277)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3575..3715
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3575..3704
                     /label=AAV2 ITR
     rep_origin      3790..4245
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3877..3896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4087..4108
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4262..4281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4381..4403
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4441..4459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4527..4631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4632..4700,4701..5492)
                     /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(4850..4869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5663..6251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6152..6171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6313..6453
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6324..6453
                     /label=AAV2 ITR
ORIGIN
        1 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
       61 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
      121 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
      181 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
      241 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
      301 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
      361 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
      421 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat
      481 tcgcggccgc caccatggga tgtatcaaat cgaaaggtaa ggacagcctg tcaggaagtg
      541 gttgcattaa gagcaaggga aaggattcgt tctgctatga aaatgaagtt gccagatctg
      601 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      661 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      721 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      781 tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
      841 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      901 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      961 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1021 tggagtacaa ctggaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     1081 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     1141 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1201 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1261 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagctcgaga
     1321 agatgaagcg gctggtgtac atcagcaaga tcagcggcca cctgagcctg gaagagatcc
     1381 agcggatcgg caaggtgtcc atcaagaaca accagcggga caacatcacc ggcgtgctgc
     1441 tgtacctgca gggcctgttc ttccagatcc tggaaggcga gaacgagaag gtggacaagc
     1501 tgtacaagaa aatcctggtg gacgaccggc acaccaacat cctgtgcctg aaaaccgagt
     1561 acgacatcac cgaccggatg ttccccaact gggccatgaa aaccatcaac ctgaacgaga
     1621 acagcgagct gatgatccag cccatcaaga gcctgctgca gaccatcacc cagagccacc
     1681 gggtgctgga aaagtacatg cccgccagag tgatctacct gatcaaccag ggcatcaacc
     1741 ccctgaccgt ggagccccag ctggtcgaga agatcatctt cttcagcgac atcctggcct
     1801 tcagcaccct gaccgagaag ctgcccgtga acgaggtggt catcctggtc aaccggtact
     1861 tcagcatctg cacccggatc atcagcgcct acggcggcga agtgaccaaa tatattggcg
     1921 attgcgtgat ggccagcttc accaaagagc agggcgacgc cgccatccgg accagcctgg
     1981 acatcatctc cgagctgaag cagctgcggc accacgtgga ggccaccaac cccctgcacc
     2041 tgctgtacac cggcatcggc ctgagctacg gccacgtgat cgagggcaac atgggcagca
     2101 gcctgaagat ggaccacacc ctgctggggg acgccgtgaa cgtggccgcc aggctggaag
     2161 ccctgacaag acagctgccc tacgccctgg cctttacagc cggcgtgaag aagtgctgcc
     2221 aggcccagtg gaccttcatc aacctgggag cccaccaggt caagggcaag caggaagcca
     2281 tcgaggtcta caccgtgaat gaggcccaga agtactacga caccctgcag atcacccagc
     2341 tgatccggca gaccctggaa aacgacaagt agactagtaa gcttatcgat aatcaacctc
     2401 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2461 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2521 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2581 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2641 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2701 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2761 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     2821 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2881 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2941 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag
     3001 acgcgatcgc ggcgcgccgg cctctctggc ctcgacctcg agtctagaga tgatctacgg
     3061 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     3121 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     3181 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     3241 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     3301 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     3361 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     3421 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     3481 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     3541 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3601 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3661 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3721 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     3781 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3841 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4081 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     4141 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4201 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4261 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4321 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4381 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4441 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4501 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5521 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6301 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     6361 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     6421 agggagtggc caactccatc actaggggtt cctgcggccg cacgcgtcga
//
