LOCUS       sequence-434847-        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  General mammalian expression of PACmn, a membrane targeted version 
            of the photoactivatable adenylyl cyclase from Beggiatoa with lowered
            dark activity, with dark Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  TITLE     cAMP in synaptic plasticity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6534)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV	nEF	2xLyn-ERex-Venus(Y145W)-bPAC(F198Y)
FEATURES             Location/Qualifiers
     source          1..6534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..244
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             257..525
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      559..568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(670..672,673..1383)
                     /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(712..733)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(973..992)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1320..1341
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2462..3050
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2515..2535)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2933..2944)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3053..3069)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3054..3070)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3137..3613
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3336..3355)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3653..3793
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3653..3782
                     /label=AAV2 ITR
     rep_origin      3868..4323
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3955..3974)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4165..4186
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4340..4359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4459..4481
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4519..4537)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4605..4709
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4710..4778,4779..5570)
                     /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(4928..4947)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5741..6329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6230..6249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6391..6520
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
       61 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
      121 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
      181 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
      241 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
      301 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
      361 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
      421 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
      481 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
      541 gcctacgaat tcgcggccgc caccatggga tgtatcaaat cgaaaggtaa ggacagcctg
      601 tcaggaagtg gttgcattaa gagcaaggga aaggattcgt tctgctatga aaatgaagtt
      661 gccagatctg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      721 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      781 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
      841 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
      901 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      961 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1021 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1081 gggcacaagc tggagtacaa ctggaacagc cacaacgtct atatcaccgc cgacaagcag
     1141 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     1201 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1261 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     1321 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1381 aagctcgaga agatgaagcg gctggtgtac atcagcaaga tcagcggcca cctgagcctg
     1441 gaagagatcc agcggatcgg caaggtgtcc atcaagaaca accagcggga caacatcacc
     1501 ggcgtgctgc tgtacctgca gggcctgttc ttccagatcc tggaaggcga gaacgagaag
     1561 gtggacaagc tgtacaagaa aatcctggtg gacgaccggc acaccaacat cctgtgcctg
     1621 aaaaccgagt acgacatcac cgaccggatg ttccccaact gggccatgaa aaccatcaac
     1681 ctgaacgaga acagcgagct gatgatccag cccatcaaga gcctgctgca gaccatcacc
     1741 cagagccacc gggtgctgga aaagtacatg cccgccagag tgatctacct gatcaaccag
     1801 ggcatcaacc ccctgaccgt ggagccccag ctggtcgaga agatcatctt cttcagcgac
     1861 atcctggcct tcagcaccct gaccgagaag ctgcccgtga acgaggtggt catcctggtc
     1921 aaccggtact tcagcatctg cacccggatc atcagcgcct acggcggcga agtgaccaaa
     1981 tatattggcg attgcgtgat ggccagcttc accaaagagc agggcgacgc cgccatccgg
     2041 accagcctgg acatcatctc cgagctgaag cagctgcggc accacgtgga ggccaccaac
     2101 cccctgcacc tgctgtacac cggcatcggc ctgagctacg gccacgtgat cgagggcaac
     2161 atgggcagca gcctgaagat ggaccacacc ctgctggggg acgccgtgaa cgtggccgcc
     2221 aggctggaag ccctgacaag acagctgccc tacgccctgg cctttacagc cggcgtgaag
     2281 aagtgctgcc aggcccagtg gaccttcatc aacctgggag cccaccaggt caagggcaag
     2341 caggaagcca tcgaggtcta caccgtgaat gaggcccaga agtactacga caccctgcag
     2401 atcacccagc tgatccggca gaccctggaa aacgacaagt agactagtta agcttatcga
     2461 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2521 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2581 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2641 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2701 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2761 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2821 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2881 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2941 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3001 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3061 tcgacctcga gacgcgatcg cggcgcgcca ttcgatggcc tctctggcct cgacctcgag
     3121 tctagagatg atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3181 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3241 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3301 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3361 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3421 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     3481 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3541 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3601 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     3661 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     3721 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     3781 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3841 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3901 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     3961 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4021 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4081 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4141 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     4201 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     4261 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4321 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4381 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4441 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4501 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4561 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4621 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4681 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4741 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4801 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4861 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4921 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4981 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5041 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5101 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5161 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5221 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5281 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5341 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5401 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5461 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5521 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5581 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5641 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5701 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5761 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5821 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5881 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5941 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6001 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6061 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6121 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6181 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6241 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6301 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6361 tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca
     6421 ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
     6481 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac gcgt
//
