LOCUS       sequence-189963-        5611 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV expression of Ruby2 spaghetti monster FLAG construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5611)
  AUTHORS   Viswanathan S, Williams ME, Bloss EB, Stasevich TJ, Speer CM, Nern 
            A, Pfeiffer BD, Hooks BM, Li WP, English BP, Tian T, Henry GL, 
            Macklin JJ, Patel R, Gerfen CR, Zhuang X, Wang Y, Rubin GM, Looger 
            LL
  TITLE     High-performance probes for light and electron microscopy.
  JOURNAL   Nat Methods. 2015 Jun;12(6):568-76. doi: 10.1038/nmeth.3365. Epub 
            2015 Apr 27.
  PUBMED    25915120
REFERENCE   2  (bases 1 to 5611)
  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: 
            "10.1038/nmeth.3365"; journalName: "Nat Methods"; date: "2015-06"; 
            volume: "12"; issue: "6"; pages: "568-76"
COMMENT     Sequence Label: pAAV.CAG.Ruby2sm-Flag.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5611
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        427..682
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
     primer_bind     961..983
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1045..1064
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1108..1117
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1117..1140
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1144..1167
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1171..1194
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1699..1722
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1729..1752
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1765..1788
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1795..1818
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2011..2034
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2038..2061
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2065..2088
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2116..2704
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2169..2189)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2587..2598)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2747..2868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2758..2777)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2812..2831
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3030..3159
                     /label=AAV2 ITR
     rep_origin      3231..3686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3318..3337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3528..3549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3712..3816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3817..3885,3886..4677)
                     /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(4035..4054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4848..5436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5337..5356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
       61 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta
      121 gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga agagtaccat
      181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      301 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      361 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      421 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      481 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      541 ggggggggcg cgcgccaggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
      601 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
      661 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
      721 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      781 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
      841 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
      901 gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
      961 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     1021 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     1081 gcaaagaatt ggatccgcgg ccgcgccgcc accatggact acaaggacga cgacgacaaa
     1141 ggtgactaca aagatgatga cgataaaggc gactataagg acgatgacga caagggcgga
     1201 aactcactga tcaaggaaaa catgcggatg aaggtggtga tggagggctc cgtgaatggt
     1261 caccagttca agtgcaccgg agagggagag ggaaacccgt acatgggaac tcagaccatg
     1321 cgcattaagg tcatcgaagg aggtccgctg ccgttcgctt tcgatatcct ggccacttcg
     1381 ttcggaggag ggtcgcgcac gttcatcaag tacccgaagg gaatcccgga cttctttaag
     1441 cagtcattcc cggaaggatt cacttgggaa cgggtgaccc ggtatgaaga tggaggtgtg
     1501 gtgactgtca tgcaagatac ttcgctggag gatgggtgcc tcgtgtacca cgtccaagtc
     1561 cgcggagtga atttcccgtc caacggacca gtgatgcaga aaaagacgaa gggttgggaa
     1621 cctaatactg aaatgatgta ccccgcagac ggagggctga ggggctacac ccacatggcg
     1681 ctgaaggtcg acggaggaga ttacaaggat gacgacgata agcaacaaga ttacaaagac
     1741 gatgatgaca agggccagca gggcgactac aaggacgacg acgacaagca gcaggactac
     1801 aaagatgacg atgataaagg aggaggacat ctgtcctgtt cgttcgtgac cacctacaga
     1861 tcaaagaaaa ccgtgggaaa catcaagatg ccgggcattc atgccgtcga ccaccgcctg
     1921 gagcggctcg aagaatcaga caatgagatg ttcgtcgtgc aaagagaaca tgccgtggcc
     1981 aagttcgcgg gactgggagg cggtggaggc gattacaaag acgatgatga caagggtgac
     2041 tataaagacg acgatgacaa aggggattac aaggatgatg atgataagta gaccggtgaa
     2101 ttcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2161 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2221 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2281 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2341 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2401 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2461 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2521 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2581 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2641 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2701 ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag atacattgat
     2761 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2821 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2881 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     2941 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     3001 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     3061 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     3121 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata gcgaagaggc
     3181 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     3241 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3301 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3361 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3421 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3481 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3541 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3601 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3661 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     3721 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3781 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3841 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3901 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3961 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4021 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4081 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4141 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4201 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4261 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4321 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4381 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4441 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4501 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4561 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4621 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4681 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4741 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4861 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4921 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4981 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5041 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5101 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5161 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5221 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5281 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5341 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5401 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5461 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5521 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5581 accgagcgca gcgagtcagt gagcgaggaa g
//
