LOCUS       sequence-471081-        5169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of a fluorescent marker, mScarletI, fused to the 
            plasma membrane targeting sequence CAAX2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  AUTHORS   Benedetti L, Fan R, Weigel AV, Moore AS, Houlihan PR, Kittisopikul 
            M, Park G, Petruncio A, Hubbard PM, Pang S, Xu CS, Hess HF, Saalfeld
            S, Rangaraju V, Clapham DE, De Camilli P, Ryan TA, 
            Lippincott-Schwartz J
  TITLE     Periodic ER-plasma membrane junctions support long-range Ca(2+) 
            signal integration in dendrites.
  JOURNAL   Cell. 2025 Jan 23;188(2):484-500.e22. doi: 
            10.1016/j.cell.2024.11.029. Epub 2024 Dec 20.
  PUBMED    39708809
REFERENCE   2  (bases 1 to 5169)
  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"; doi: 
            "10.1016/j.cell.2024.11.029"; journalName: "Cell"; date: 
            "2025-01-23- 23"; volume: "188"; issue: "2"; pages: "484-500.e22"
COMMENT     Sequence Label: pAAV hSyn INTRON mScarlet-CAAX2 WPRE
FEATURES             Location/Qualifiers
     source          1..5169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(996..1787,1788..1856))
                     /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     1619..1638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1857..1961)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1987..2442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2124..2145)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2336..2355
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2514..2643
                     /label=AAV2 ITR
     promoter        2725..3172
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     intron          3194..3326
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3309..3328
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3340..3349
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3346..4041
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(3479..3497)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3747..3766
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4120..4708
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4173..4193)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4591..4602)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4711..4727)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4712..4728)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4757..4878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4768..4787)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4822..4841
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5040..5169
                     /label=AAV2 ITR
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1981 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2041 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2101 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2161 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2221 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2281 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2341 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2401 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     2461 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
     2521 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     2581 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2641 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     2701 tctctgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga tgaggcgggg
     2761 tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc
     2821 ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg
     2881 cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca
     2941 ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct
     3001 tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag
     3061 gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct
     3121 gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc agtcgaattc
     3181 aagctgctag caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     3241 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     3301 catccacttt gcctttctct ccacaggtga taaggatccg ccaccatggt gagcaagggc
     3361 gaggcagtga tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc
     3421 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     3481 aagctgaagg tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag
     3541 ttcatgtacg gctccagggc cttcatcaag caccccgccg acatccccga ctactataag
     3601 cagtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc
     3661 gtgaccgtga cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc
     3721 cgcggcacca acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa
     3781 gcgtccaccg agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc
     3841 ctgcgcctga aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag
     3901 aagcccgtgc agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac
     3961 aacgaggact acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc
     4021 ggcatggacg agctgtacaa gggcggctcc ggaggcagca agaaaaagaa gaagaagtcc
     4081 aagacaaaat gcgtgattat gtagtaaaag cttatcgata atcaacctct ggattacaaa
     4141 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4201 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4261 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4321 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4381 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4441 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4501 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     4561 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4621 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4681 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg
     4741 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4801 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4861 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     4921 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     4981 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     5041 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     5101 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     5161 agcgcgcag
//
