LOCUS       sequence-201302-        7971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  - humanized - variant of pAAV hSyn YFP-CaRhAC Addgene # 101721– less
            reliable in hippocampal neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  AUTHORS   Scheib U, Broser M, Constantin OM, Yang S, Gao S, Mukherjee S, 
            Stehfest K, Nagel G, Gee CE, Hegemann P
  TITLE     Rhodopsin-cyclases for photocontrol of cGMP/cAMP and 2.3 A structure
            of the adenylyl cyclase domain.
  JOURNAL   Nat Commun. 2018 May 24;9(1):2046. doi: 10.1038/s41467-018-04428-w.
  PUBMED    29799525
REFERENCE   2  (bases 1 to 7971)
  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: "Nat 
            Commun."; date: "2018-05-24"; volume: "9(1):2046. doi"; pages: " 
            10.1038/s41467-018-04428-w"
COMMENT     Sequence Label: pAAV hSyn1 CaRhAC T2A tDimer
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   363..492
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(554..1142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(634..653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1313..2104,2105..2173))
                     /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     1936..1955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2174..2278)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2346..2364
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2402..2424)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2524..2543
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2560..3015)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2697..2718)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2909..2928
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3090..3230
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3101..3230
                     /label=AAV2 ITR
     polyA_signal    3270..3746
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3528..3547
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     3813..3829
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3814..3830
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3833..4421
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3939..3950
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4348..4368
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4449..5123)
                     /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(5166..5843)
                     /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      5836..5845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5840..5849
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5865..5918)
                     /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"
     promoter        complement(join(7838..7971,1..314))
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 ggccggcggc ggcgcggacg cgaccaaggt ggccgggaag gggagtttgc gggggaccgg
       61 cgagtgacgt cagcgcgcct tcagtgctga ggcggcggtg gcgcgcgccg ccaggcgggg
      121 gcgaaggcac tgtccgcggt gctgaagctg gcagtgcgca cgcgcctcgc cgcatcctgt
      181 ttcccctccc cctctctgat aggggatgcg caatttgggg aatgggggtt gggtgcttgt
      241 ccagtgggtc ggggtcggtc gtcaggtagg cacccccacc ccgcctcatc ctggtcctaa
      301 aacccacttg cactcatacg cagggccctc tgcagctcga gtctagcgac gcgtgcggcc
      361 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
      421 ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca
      481 gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      541 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      601 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      661 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      721 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      781 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      841 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      901 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      961 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1021 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1081 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1141 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1201 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1261 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1321 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1381 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1441 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1501 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1561 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1621 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1681 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1741 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1801 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1861 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1921 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1981 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2041 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2101 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2161 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2221 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2281 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     2341 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2401 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2461 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     2521 atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt
     2581 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa
     2641 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag
     2701 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2761 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
     2821 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     2881 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg
     2941 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3001 cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg
     3061 cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac
     3121 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     3181 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgctc
     3241 ggtccgcacg tggttaccta caaaatcaga aggacaggga agggagcagt ggttcacgcc
     3301 tgtaatccca gcaatttggg aggccaaggt gggtagatca cctgagatta ggagttggag
     3361 accagcctgg ccaatatggt gaaaccccgt ctctaccaaa aaaacaaaaa ttagctgagc
     3421 ctggtcatgc atgcctggaa tcccaacaac tcgggaggct gaggcaggag aatcgcttga
     3481 acccaggagg cggagattgc agtgagccaa gattgtgcca ctgcactcca gcttggttcc
     3541 caatagaccc cgcaggccct acaggttgtc ttcccaactt gccccttgct ccataccacc
     3601 cccctccacc ccataatatt atagaaggac acctagtcag acaaaatgat gcaacttaat
     3661 tttattagga caaggctggt gggcactgga gtggcaactt ccagggccag gagaggcact
     3721 ggggaggggt cacagggatg ccacccgtag atcatctcta gactcgaggt cgaggccaga
     3781 gaggccatcg aatggcgcgc cgcgatcgcg tctcgaggtc gacggtatcg atgcggggag
     3841 gcggcccaaa gggagatccg actcgtctga gggcgaaggc gaagacgcgg aagaggccgc
     3901 agagccggca gcaggccgcg ggaaggaagg tccgctggat tgagggccga agggacgtag
     3961 cagaaggacg tcccgcgcag aatccaggtg gcaacacagg cgagcagcca aggaaaggac
     4021 gatgatttcc ccgacaacac cacggaattg tcagtgccca acagccgagc ccctgtccag
     4081 cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa tagggagggg gaaagcgaaa
     4141 gtcccggaaa ggagctgaca ggtggtggca atgccccaac cagtgggggt tgcgtcagca
     4201 aacacagtgc acaccacgcc acgttgcctg acaacgggcc acaactcctc ataaagagac
     4261 agcaaccagg atttatacaa ggaggagaaa atgaaagcca tacgggaagc aatagcatga
     4321 tacaaaggca ttaaagcagc gtatccacat agcgtaaaag gagcaacata gttaagaata
     4381 ccagtcaatc tttcacaaat tttgtaatcc agaggttgat tatcgataag cttttgctcg
     4441 aagtcgacct acaggaacag gtggtggcgg ccctcggagc gctcgtactg ttccacgatg
     4501 gtgtagtcct cgttgtggga ggtgatgtcc agcttggtgt ccacgtagta gtagccgggc
     4561 agctgcacgg gcttcttggc catgtagatg gtcttgaact ccaccaggta gtggccgccg
     4621 tccttcagct tcagggcctg gtggatctcg cccttcagca cgccgtcgcg ggggtacagg
     4681 cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggggggg
     4741 aagttggtgc cgcggaactt caccttgtag atcagcgtgc cgtcctgcag ggaggagtcc
     4801 tgggtcacgg tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc
     4861 tcggggaagg acagcttctt gtagtcgggg atgtcggcgg ggtgcttcac gtacgccttg
     4921 gagccgtact ggaactgggg ggacaggatg tcccaggcga agggcagggg gccgcccttg
     4981 gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg
     5041 atctcgaact cgtggccgtt cacggagccc tccatgcgca ccttgaagcg catgaactct
     5101 ttgatgacgt cctcggagga ggcggtgccg gagctgccgc tgccggtgct gccggtgcca
     5161 tgccccagga acaggtggtg gcggccctcg gagcgctcgt actgttccac gatggtgtag
     5221 tcctcgttgt gggaggtgat gtccagcttg gtgtccacgt agtagtagcc gggcagctgc
     5281 acgggcttct tggccatgta gatggtcttg aactccacca ggtagtggcc gccgtccttc
     5341 agcttcaggg cctggtggat ctcgcccttc agcacgccgt cgcgggggta caggcgctcg
     5401 gtggaggcct cccagcccat ggtcttcttc tgcattacgg ggccgtcggg ggggaagttg
     5461 gtgccgcgga acttcacctt gtagatcagc gtgccgtcct gcagggagga gtcctgggtc
     5521 acggtcacca cgccgccgtc ctcgaagttc atcacgcgct cccacttgaa gccctcgggg
     5581 aaggacagct tcttgtagtc ggggatgtcg gcggggtgct tcacgtacgc cttggagccg
     5641 tactggaact ggggggacag gatgtcccag gcgaagggca gggggccgcc cttggtcacc
     5701 ttcagcttgg cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg
     5761 aactcgtggc cgttcacgga gccctccatg cgcaccttga agcgcatgaa ctctttgatg
     5821 acgtcctcgg aggaggccac catggcggcc gcggatcccg gtgcagggcc gggattctcc
     5881 tccacgtcac cgcatgttag aagacttcct ctgccctcgg ccgcgggtac cgactttcta
     5941 gcggtcaccc aatatgtctg catttttccc tttcctttca cttccatcag gtccagtggc
     6001 tgggtcacga attttccctt gatcatctgg taagtagagt cggagatatg aatgtgccct
     6061 gccttgctgg ttgactccat ccgggaagct gtattgacgg tatcccccac cagatcccaa
     6121 tgagggttca ggtcgcccag gactccggct gtcactggtc cactattcag gccgatgcga
     6181 atgttgatag attcgccggt agcagtttta aaagtcttga tcatctcgat gatgtccagg
     6241 gcgaagttca ctgctcgatc agcgtggtcg gggaccacct ctggtgctcc ggtcactccc
     6301 agataagcgt ccccaatagt cttgacctta tacacgcccc accgcttagc gatggcatca
     6361 tactccagcc acagcttgtt cagggtggcc atcatgtcct ttgtggaggt cctggagcta
     6421 atcacagtaa agtttgtgat gtcagaaaag aagactgtca cggattcgta ctctttggcc
     6481 tcggttgcct gtccattctt cagctgttcc agcaccccct caggcatcat cttctgcaga
     6541 atggcgtcac agttagtgat ctgagattcc agttcctctg caatagctgt gatggcttcg
     6601 actttctgat tctgtgcctg ctccacagtg ctgttgacca gcaccagtgt caggaacacc
     6661 ttggccccaa tatcgatcat ggtaaaagat gcctcagtca ttgtgaaact cagcatgcca
     6721 gatgtaaagg cgaaccagct cagtggaaac agggtccatg tggtgacagt tgagaagcgt
     6781 gtccacctca ggccattcac ttcaatgttg caaggggtct ctccgttgat ggccccagta
     6841 aaggacctcc acaggctgta gaccacgaag ctaaagaagg cacatgacag gatattaaac
     6901 aggtttcccc agggataagg gggcagcaca gctccaaaga acccgcagac caccagggcg
     6961 taatcagaga agaccacccc ccaggcatta tggtctgcct gtgtaatctc ggagatcagc
     7021 agaatcagga cgggacaggt agcaatccac tccaggtatc ttgcaggctc cactgggttg
     7081 ctcagggtgc cttcgatggt tggagtcagc cgcagcataa tcaggatgta tgaactaaaa
     7141 gcgatgatgt tcactcccag caggcacagg gacagtgctt tcttctctgg gtttctggcg
     7201 cctgcaaatc ccatataact gaaagcaatc ccgatgccaa atccgaaggc cccatagccg
     7261 taccaaatgt ctgcccagcc cctattataa gcctgtcttt cctggataaa cagagcgtag
     7321 aaagcgccgg ccacggtcag aatggcccac atgataaact gtcctgtcag gaatttcagt
     7381 gaaggggtac tcagcatcat atcccaggac cacatgtctg cgatgtttgt ctggtactgc
     7441 tgcagagcgg actgcttact aggagagcca ggtcggctat tggcgggaga ggacacgttg
     7501 cttgaacttg cagaggaggt gctgcggaca ttggacacgc tcagtgaatt gttctgtctg
     7561 ttccgcatat tagcgttgct tgaggcggta ctagagccag tgaccttgat attgggctga
     7621 ctgggagatg gcagcagtga ggctccggag ctgcggaaac ttccgtttcg agtcagtggc
     7681 tgctcgacga tagggtgttt tgtcacgcta cacttgcaat cgtcgcacag ggttgaggtt
     7741 ggtgagtaac agtattcagg gcaggaacag ccactacatg ctccccgcag gttattatct
     7801 ttatctttca tagacataga cattattgcg aattcgactg cgctctcagg cacgacacga
     7861 ctcctccgct gcccaccgca gactgaggca gcgctgagtc gccggcgccg cagcgcagat
     7921 ggtcgcgccc gtgcccccct atctcgcgcc tcgcgtggtg cggtccggct g
//
