LOCUS       sequence-409232-        7452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid for CaMKIIa driven co-expression of mScarlet-I and 
            2M-Qt-FLAG EMcapsulins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7452)
  AUTHORS   Sigmund F, Berezin O, Beliakova S, Magerl B, Drawitsch M, Piovesan 
            A, Goncalves F, Bodea SV, Winkler S, Bousraou Z, Grosshauser M, 
            Samara E, Pujol-Marti J, Schadler S, So C, Irsen S, Walch A, Kofler 
            F, Piraud M, Kornfeld J, Briggman K, Westmeyer GG
  TITLE     Genetically encoded barcodes for correlative volume electron 
            microscopy.
  JOURNAL   Nat Biotechnol. 2023 Apr 17. doi: 10.1038/s41587-023-01713-y.
  PUBMED    37069313
REFERENCE   2  (bases 1 to 7452)
  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 
            Biotechnol. 2023 Apr 17. doi: 10.1038/s41587-023-01713-y."
COMMENT     Sequence Label: CaMKIIa-mScarlet-I-P2A-2M-Qt-FLAG
FEATURES             Location/Qualifiers
     source          1..7452
                     /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        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1452..1461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2163..2219
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3528..3551
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3579..4167
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3632..3652)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4050..4061)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4199..4675
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4398..4417)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4715..4855
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4715..4844
                     /label=AAV2 ITR
     rep_origin      4930..5385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5017..5036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5227..5248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5402..5421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5521..5543
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5581..5599)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5667..5771
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5772..5840,5841..6632)
                     /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(5990..6009)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6803..7391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7292..7311
                     /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cgccaccatg gtttctaaag gcgaggccgt gatcaaagag ttcatgaggt
     1501 tcaaggtgca catggaaggc agcatgaacg gccacgagtt cgagatcgaa ggcgaaggcg
     1561 agggcagacc ttacgaggga actcagaccg ccaagctgaa agtgaccaaa ggcggccctc
     1621 tgcctttcag ctgggacatc ctgtctccac agtttatgta cggcagcagg gccttcatca
     1681 agcaccccgc tgacatcccc gactactaca agcagtcttt cccagagggc ttcaagtggg
     1741 agagagtgat gaacttcgag gacggcggag ccgtgactgt gacacaggac acatctctgg
     1801 aagatgggac cctgatctac aaagtgaagc tgcggggcac aaacttccct ccagacggcc
     1861 ctgtgatgca gaaaaagaca atgggctggg aagccagcac cgagagactg taccctgaag
     1921 atggcgtgct gaagggcgac atcaagatgg ccctgagact gaaggatggc ggcagatacc
     1981 tggccgactt caagaccacc tacaaggcca agaaacctgt gcagatgcct ggcgcctaca
     2041 acgtggacag aaagctggac atcaccagcc acaacgagga ctacaccgtg gtggaacagt
     2101 acgagagaag cgaggggaga cacagcacag gcggaatgga cgagctgtac aagggctctg
     2161 gcgccacaaa cttcagcctg cttaaacagg ccggcgacgt ggaagagaac cccggaccta
     2221 tggacccaga gacctgtccg tgtccaactg gaggttcttg tacgtgtagt gataagtgta
     2281 aatgcaaggg atgtaaatgt actaactgta agaaatcatg ttgtagttgt tgccctgccg
     2341 gatgtgagaa atgtgcgaaa gactgtgttt gtaaggggga agagggtgcc aaagctgagg
     2401 cggaaaaatg ttcctgttgt caaggtggag gttctggcgg tggatcggga ggttcagcgg
     2461 atcctgaaac atgtccttgt cctactggtg gaagttgtac ttgttctgat aaatgtaagt
     2521 gcaaaggctg caaatgtacc aattgtaaaa aatcttgttg ttcgtgttgt cccgctggtt
     2581 gtgaaaagtg tgctaaagat tgtgtatgta aaggtgaaga aggagcaaaa gcagaagccg
     2641 agaagtgttc atgctgtcag ggcagcggcc cgggcaacaa gagccagctg taccctgaca
     2701 gccctctgac cgaccaggac ttcaaccagc tggaccagac cgtgattgaa gccgccagaa
     2761 ggcagctcgt cggcagaaga ttcatcgagc tgtacggccc tctcggcaga ggcatgcaga
     2821 gcgtgttcaa cgacatcttc atggaaagcc acgaggccaa gatggacttc cagggcagct
     2881 tcgacaccga ggtggaaagc agcagaagag tgaactacac aatccccatg ctgtacaagg
     2941 acttcgtgct gtactggcgc gacctggaac agagcaaggc cctggacatc cccatcgatt
     3001 tttccgtggc cgccaacgcc gctagagatg tggcctttct ggaagatcag atgatctttc
     3061 acggcagcaa agagttcgac atccccggcc tgatgaacgt gaagggcaga ctgacacacc
     3121 tgatcggcaa ttggtacgag agcggcaacg ccttccagga catcgtggaa gcccggaaca
     3181 agctgctgga aatgaaccac aacggcccct acgctctggt gctgagccct gagctgtata
     3241 gcctgctgca cagagtgcac aaggacacca acgtcctgga aatcgagcac gtgcgcgagc
     3301 tgattaccgc cggtgtcttt cagagccctg tgctgaaggg aaagtccggc gtgatcgtga
     3361 acaccggcag aaacaacctg gacctggcca tcagcgagga cttcgagaca gcctatctgg
     3421 gcgaagaggg catgaatcac cccttccggg tgtacgagac agtggtgctg cggattaaga
     3481 ggcctgccgc catctgcacc ctgatcgatc ctgaagaagg ctccggcgac tacaaggacg
     3541 acgacgacaa atgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     3601 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3661 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3721 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3781 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3841 gtcagctcct ttccggaact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3901 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3961 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
     4021 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     4081 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     4141 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg
     4201 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     4261 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     4321 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     4381 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     4441 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     4501 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     4561 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     4621 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     4681 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4741 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4801 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4861 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4921 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4981 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     5041 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5101 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5161 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5221 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     5281 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     5341 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5401 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5461 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5521 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5581 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5641 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5701 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5761 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5821 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5881 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5941 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6001 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6061 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6121 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6181 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6241 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6301 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6361 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6421 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6481 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6541 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6601 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6661 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6721 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6781 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6841 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6901 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6961 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7021 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7081 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7141 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7201 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7261 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7321 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7381 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7441 ttgctcacat gt
//
