LOCUS       sequence-434848-        6400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuronal expression of PACmn with mKate2, a membrane targeted 
            version of the photoactivatable adenylyl cyclase from Beggiatoa with
            lowered dark activity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6400)
  TITLE     cAMP in synaptic plasticity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6400)
  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            "Unpublished"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 Syn 2xLyn-ERex-mKate2-bPAC(F198Y)
FEATURES             Location/Qualifiers
     source          1..6400
                     /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        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      642..651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             753..1448
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    2527..3115
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2580..2600)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2998..3009)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3147..3623
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3346..3365)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3663..3803
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3663..3792
                     /label=AAV2 ITR
     rep_origin      3878..4333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3965..3984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4175..4196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4350..4369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4469..4491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4529..4547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4615..4719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4720..4788,4789..5580)
                     /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(4938..4957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5751..6339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6240..6259
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc cgccaccatg ggatgtatca
      661 aatcgaaagg taaggacagc ctgtcaggaa gtggttgcat taagagcaag ggaaaggatt
      721 cgttctgcta tgaaaatgaa gttgccagat ctatggtgag cgagctgatt aaggagaaca
      781 tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg
      841 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggcg gtcgagggcg
      901 gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc agcaaaacct
      961 tcatcaacca cacccagggc atccccgact tctttaagca gtccttcccc gagggcttca
     1021 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     1081 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca
     1141 acggccctgt gatgcagaag aaaacactcg gctgggaggc ctccaccgag accctgtacc
     1201 ccgctgacgg cggcctggaa ggcagagccg acatggccct gaagctcgtg ggcgggggcc
     1261 acctgatctg caacttgaag accacataca gatccaagaa acccgctaag aacctcaaga
     1321 tgcccggcgt ctactatgtg gacagaagac tggaaagaat caaggaggcc gacaaagaga
     1381 cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg
     1441 ggcacagact cgagaagatg aagcggctgg tgtacatcag caagatcagc ggccacctga
     1501 gcctggaaga gatccagcgg atcggcaagg tgtccatcaa gaacaaccag cgggacaaca
     1561 tcaccggcgt gctgctgtac ctgcagggcc tgttcttcca gatcctggaa ggcgagaacg
     1621 agaaggtgga caagctgtac aagaaaatcc tggtggacga ccggcacacc aacatcctgt
     1681 gcctgaaaac cgagtacgac atcaccgacc ggatgttccc caactgggcc atgaaaacca
     1741 tcaacctgaa cgagaacagc gagctgatga tccagcccat caagagcctg ctgcagacca
     1801 tcacccagag ccaccgggtg ctggaaaagt acatgcccgc cagagtgatc tacctgatca
     1861 accagggcat caaccccctg accgtggagc cccagctggt cgagaagatc atcttcttca
     1921 gcgacatcct ggccttcagc accctgaccg agaagctgcc cgtgaacgag gtggtcatcc
     1981 tggtcaaccg gtacttcagc atctgcaccc ggatcatcag cgcctacggc ggcgaagtga
     2041 ccaaatatat tggcgattgc gtgatggcca gcttcaccaa agagcagggc gacgccgcca
     2101 tccggaccag cctggacatc atctccgagc tgaagcagct gcggcaccac gtggaggcca
     2161 ccaaccccct gcacctgctg tacaccggca tcggcctgag ctacggccac gtgatcgagg
     2221 gcaacatggg cagcagcctg aagatggacc acaccctgct gggggacgcc gtgaacgtgg
     2281 ccgccaggct ggaagccctg acaagacagc tgccctacgc cctggccttt acagccggcg
     2341 tgaagaagtg ctgccaggcc cagtggacct tcatcaacct gggagcccac caggtcaagg
     2401 gcaagcagga agccatcgag gtctacaccg tgaatgaggc ccagaagtac tacgacaccc
     2461 tgcagatcac ccagctgatc cggcagaccc tggaaaacga caagtagact agttaagctt
     2521 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2581 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2641 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2701 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2761 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2821 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2881 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     2941 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3001 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3061 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     3121 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
     3181 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     3241 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     3301 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     3361 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     3421 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     3481 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     3541 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     3601 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     3661 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3721 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3781 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3841 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3901 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3961 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4021 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4081 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4141 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4201 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     4261 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4321 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4381 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4441 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4501 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4561 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4621 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4681 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4741 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4801 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4861 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4921 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4981 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5041 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5101 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5161 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5221 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5281 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5341 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5401 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5461 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5521 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5581 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5881 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6361 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
