LOCUS       sequence-330311-        6472 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 6472)
  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 6472)
  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..6472
                     /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        179..626
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      641..650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(752..754,755..1465)
                     /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(794..815)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1055..1074)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1402..1423
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2544..3132
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2597..2617)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3015..3026)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3135..3151)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3136..3152)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3219..3695
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3418..3437)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3735..3875
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3735..3864
                     /label=AAV2 ITR
     rep_origin      3950..4405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4037..4056)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4247..4268
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4422..4441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4541..4563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4601..4619)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4687..4791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4792..4860,4861..5652)
                     /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(5010..5029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5823..6411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6312..6331
                     /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 gcgtcgctag actcgagctg cagagggccc tgcgtatgag
      181 tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg acgaccgacc
      241 ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga
      301 gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga
      361 cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc
      421 tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg
      481 ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg
      541 accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag
      601 gagtcgtgtc gtgcctgaga gcgcagtcga attcgcggcc gccaccatgg gatgtatcaa
      661 atcgaaaggt aaggacagcc tgtcaggaag tggttgcatt aagagcaagg gaaaggattc
      721 gttctgctat gaaaatgaag ttgccagatc tgtgagcaag ggcgaggagc tgttcaccgg
      781 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      841 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
      901 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg
      961 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1021 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1081 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1141 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactggaaca gccacaacgt
     1201 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     1261 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1321 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     1381 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1441 tctcggcatg gacgagctgt acaagctcga gaagatgaag cggctggtgt acatcagcaa
     1501 gatcagcggc cacctgagcc tggaagagat ccagcggatc ggcaaggtgt ccatcaagaa
     1561 caaccagcgg gacaacatca ccggcgtgct gctgtacctg cagggcctgt tcttccagat
     1621 cctggaaggc gagaacgaga aggtggacaa gctgtacaag aaaatcctgg tggacgaccg
     1681 gcacaccaac atcctgtgcc tgaaaaccga gtacgacatc accgaccgga tgttccccaa
     1741 ctgggccatg aaaaccatca acctgaacga gaacagcgag ctgatgatcc agcccatcaa
     1801 gagcctgctg cagaccatca cccagagcca ccgggtgctg gaaaagtaca tgcccgccag
     1861 agtgatctac ctgatcaacc agggcatcaa ccccctgacc gtggagcccc agctggtcga
     1921 gaagatcatc ttcttcagcg acatcctggc cttcagcacc ctgaccgaga agctgcccgt
     1981 gaacgaggtg gtcatcctgg tcaaccggta cttcagcatc tgcacccgga tcatcagcgc
     2041 ctacggcggc gaagtgacca aatatattgg cgattgcgtg atggccagct tcaccaaaga
     2101 gcagggcgac gccgccatcc ggaccagcct ggacatcatc tccgagctga agcagctgcg
     2161 gcaccacgtg gaggccacca accccctgca cctgctgtac accggcatcg gcctgagcta
     2221 cggccacgtg atcgagggca acatgggcag cagcctgaag atggaccaca ccctgctggg
     2281 ggacgccgtg aacgtggccg ccaggctgga agccctgaca agacagctgc cctacgccct
     2341 ggcctttaca gccggcgtga agaagtgctg ccaggcccag tggaccttca tcaacctggg
     2401 agcccaccag gtcaagggca agcaggaagc catcgaggtc tacaccgtga atgaggccca
     2461 gaagtactac gacaccctgc agatcaccca gctgatccgg cagaccctgg aaaacgacaa
     2521 gtagactagt taagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2581 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2641 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2701 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2761 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2821 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2881 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2941 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3001 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3061 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3121 ccgcctcccc gcatcgatac cgtcgacctc gagacgcgat cgcggcgcgc cattcgatgg
     3181 cctctctggc ctcgacctcg agtctagaga tgatctacgg gtggcatccc tgtgacccct
     3241 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     3301 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     3361 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     3421 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     3481 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     3541 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     3601 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     3661 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     3721 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     3781 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     3841 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     3901 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     3961 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4021 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4081 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4141 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     4201 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4261 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     4321 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4381 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     4441 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4501 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4561 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     4621 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4681 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4741 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4801 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5641 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5941 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
