LOCUS       sequence-201308-        7170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  humanized photoactivated adenylyl cyclase lower dark activity, 
            spectrum shifted 15 nm longer wavelenght with neuron-specific 
            promoter, plus red fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7170)
  TITLE     Thomas Oertner Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7170)
  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            "Unpublished"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 hSyn1 bPAC cMyc S27A 2A tDimer
FEATURES             Location/Qualifiers
     source          1..7170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   380..509
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(571..1159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(651..670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1330..2121,2122..2190))
                     /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     1953..1972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2191..2295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2363..2381
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2419..2441)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2541..2560
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2577..3032)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2714..2735)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2926..2945
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3107..3247
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3118..3247
                     /label=AAV2 ITR
     polyA_signal    3287..3763
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3545..3564
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     3830..3846
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3831..3847
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3850..4438
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3956..3967
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4365..4385
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4466..5140)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     CDS             complement(5183..5860)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     regulatory      5853..5862
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5857..5866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5882..5935)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(5948..5977)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(join(7054..7170,1..331))
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 gtggtgcggt ccggctgggc cggcggcggc gcggacgcga ccaaggtggc cgggaagggg
       61 agtttgcggg ggaccggcga gtgacgtcag cgcgccttca gtgctgaggc ggcggtggcg
      121 cgcgccgcca ggcgggggcg aaggcactgt ccgcggtgct gaagctggca gtgcgcacgc
      181 gcctcgccgc atcctgtttc ccctccccct ctctgatagg ggatgcgcaa tttggggaat
      241 gggggttggg tgcttgtcca gtgggtcggg gtcggtcgtc aggtaggcac ccccaccccg
      301 cctcatcctg gtcctaaaac ccacttgcac tcatacgcag ggccctctgc agctcgagtc
      361 tagcgacgcg tgcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
      421 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt
      481 gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc aaaaggccag
      541 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      601 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      661 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      721 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      781 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      841 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      901 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      961 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1021 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1081 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1321 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1381 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1441 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1501 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1561 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1621 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1681 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1741 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1801 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1861 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1921 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1981 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2041 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2101 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2161 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2221 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2281 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     2341 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     2401 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2461 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2521 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat
     2581 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2641 taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagataga
     2701 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2761 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     2821 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2881 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2941 aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3001 cgccgcgctt aatgcgccgc tacagggcgc ttactatggt tgctttgacg tatgcggtgt
     3061 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg caggcagctg
     3121 cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt
     3181 cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg
     3241 ggttcctgcg gccgctcggt ccgcacgtgg ttacctacaa aatcagaagg acagggaagg
     3301 gagcagtggt tcacgcctgt aatcccagca atttgggagg ccaaggtggg tagatcacct
     3361 gagattagga gttggagacc agcctggcca atatggtgaa accccgtctc taccaaaaaa
     3421 acaaaaatta gctgagcctg gtcatgcatg cctggaatcc caacaactcg ggaggctgag
     3481 gcaggagaat cgcttgaacc caggaggcgg agattgcagt gagccaagat tgtgccactg
     3541 cactccagct tggttcccaa tagaccccgc aggccctaca ggttgtcttc ccaacttgcc
     3601 ccttgctcca taccaccccc ctccacccca taatattata gaaggacacc tagtcagaca
     3661 aaatgatgca acttaatttt attaggacaa ggctggtggg cactggagtg gcaacttcca
     3721 gggccaggag aggcactggg gaggggtcac agggatgcca cccgtagatc atctctagac
     3781 tcgaggtcga ggccagagag gccatcgaat ggcgcgccgc gatcgcgtct cgaggtcgac
     3841 ggtatcgatg cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa
     3901 gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga
     3961 gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga
     4021 gcagccaagg aaaggacgat gatttccccg acaacaccac ggaattgtca gtgcccaaca
     4081 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag
     4141 ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag
     4201 tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca
     4261 actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac
     4321 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     4381 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat
     4441 cgataagctt ttgctcgaag tcgacctaca ggaacaggtg gtggcggccc tcggagcgct
     4501 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttggtgtcca
     4561 cgtagtagta gccgggcagc tgcacgggct tcttggccat gtagatggtc ttgaactcca
     4621 ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg gatctcgccc ttcagcacgc
     4681 cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc catggtcttc ttctgcatta
     4741 cggggccgtc gggggggaag ttggtgccgc ggaacttcac cttgtagatc agcgtgccgt
     4801 cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc
     4861 gctcccactt gaagccctcg gggaaggaca gcttcttgta gtcggggatg tcggcggggt
     4921 gcttcacgta cgccttggag ccgtactgga actgggggga caggatgtcc caggcgaagg
     4981 gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     5041 cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgcgcacct
     5101 tgaagcgcat gaactctttg atgacgtcct cggaggaggc ggtgccggag ctgccgctgc
     5161 cggtgctgcc ggtgccatgc cccaggaaca ggtggtggcg gccctcggag cgctcgtact
     5221 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc cagcttggtg tccacgtagt
     5281 agtagccggg cagctgcacg ggcttcttgg ccatgtagat ggtcttgaac tccaccaggt
     5341 agtggccgcc gtccttcagc ttcagggcct ggtggatctc gcccttcagc acgccgtcgc
     5401 gggggtacag gcgctcggtg gaggcctccc agcccatggt cttcttctgc attacggggc
     5461 cgtcgggggg gaagttggtg ccgcggaact tcaccttgta gatcagcgtg ccgtcctgca
     5521 gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc
     5581 acttgaagcc ctcggggaag gacagcttct tgtagtcggg gatgtcggcg gggtgcttca
     5641 cgtacgcctt ggagccgtac tggaactggg gggacaggat gtcccaggcg aagggcaggg
     5701 ggccgccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
     5761 cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgcgc accttgaagc
     5821 gcatgaactc tttgatgacg tcctcggagg aggccaccat ggcggccgcg gatcccggtg
     5881 cagggccggg attctcctcc acgtcaccgc atgttagaag acttcctctg ccctcggccg
     5941 cgggtaccag gtcctcctcc gagatcagct tttgctcgta cgtccgcggc ttgtcgtttt
     6001 ccagggtctg ccggatcagc tgggtgatct gcagggtgtc gtagtacttc tgggcctcat
     6061 tcacggtgta gacctcgatg gcttcctgct tgcccttgac ctggtgggct cccaggttga
     6121 tgaaggtcca ctgggcctgg cagcacttct tcacgccggc tgtaaaggcc agggcgtagg
     6181 gcagctgtct tgtcagggct tccagcctgg cggccacgtt cacggcgtcc cccagcaggg
     6241 tgtggtccat cttcaggctg ctgcccatgt tgccctcgat cacgtggccg tagctcaggc
     6301 cgatgccggt gtacagcagg tgcagggggt tggtggcctc cacgtggtgc cgcagctgct
     6361 tcagctcgga gatgatgtcc aggctggtcc ggatggcggc gtcgccctgc tctttggtga
     6421 agctggccat cacgcagtcg ccgatgaact tggtcacttc gccgccgtag gcgctgatga
     6481 tccgggtgca gatgctgaag taccggttga ccaggatgac cacctcgttc acgggcagct
     6541 tctcggtcag ggtgctgaag gccaggatgt cgctgaagaa gatgatcttc tcgaccagct
     6601 ggggctccac ggtcaggggg ttgatgccct ggttgatcag gtagatcact ctggcgggca
     6661 tgtacttttc cagcacccgg tggctctggg tgatggtctg cagcaggctc ttgatgggct
     6721 ggatcatcag ctcgctgttc tcgttcaggt tgatggtttt catggcccag ttggggaaca
     6781 tccggtcggt gatgtcgtac tcggttttca ggcacaggat gttggtgtgc cggtcgtcca
     6841 ccaggatttt cttgtacagc ttgtccacct tctcgttctc gccttccagg atctggaaga
     6901 acaggccctg caggtacagc agcacgccgg tgatgttgtc ccgctggttg ttcttgatgg
     6961 ccaccttgcc gatccgctgg atctcttcca ggctcaggtg gccgctgatc ttgctgatgt
     7021 acaccagccg cttcatcatg gtggcgaatt cgactgcgct ctcaggcacg acacgactcc
     7081 tccgctgccc accgcagact gaggcagcgc tgagtcgccg gcgccgcagc gcagatggtc
     7141 gcgcccgtgc ccccctatct cgcgcctcgc
//
