LOCUS       sequence-193971-        5896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of GFP spaghetti monster myc construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5896)
  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 5896)
  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.GFPsm-myc.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   3..132
                     /label=AAV2 ITR
     promoter        443..720
                     /label=chicken beta-actin promoter
     primer_bind     999..1021
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1083..1102
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1138..1147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1147..1176
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1180..1209
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1213..1242
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1774..1803
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1810..1839
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1852..1881
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1888..1917
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2131..2160
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2164..2193
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2197..2226
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2246..2834
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2299..2319)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2717..2728)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2877..2998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2888..2907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2942..2961
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3544..4132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3624..3643)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4303..5094,5095..5163))
                     /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     4926..4945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5164..5268)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5294..5749)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5431..5452)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5643..5662
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
       61 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
      121 ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg tagccatgct
      181 ctaggaagag taccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      421 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      481 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      541 atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg sgrggggsgg
      601 ggckkgkygm ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      661 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      721 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      781 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      841 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      901 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gctgtccgcg
      961 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1021 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1081 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattgg atcggccgcc
     1141 accatggagc aaaaactgat atcggaggaa gacttggcag agcagaaact gatttcggaa
     1201 gaggatcttg ccgaacaaaa actgatcagt gaggaagacc ttgccggagt gtcgaagggt
     1261 gaagagctgt tcaccggcgt ggtgcccatc ctcgtcgagc tggacggaga tgtcaatgga
     1321 cataaattct ccgtacgggg agagggcgag ggcgacgcaa ccaacggtaa actgaccctg
     1381 aagttcattt gcaccacggg taaactgccc gtaccctggc ccaccctggt cacaaccttg
     1441 ggtggcggag ttcagtgttt ttcgcggtat ccagatcaca tgaagcagca tgactttttc
     1501 aagagcgcta tgcccgaggg ttacgttcaa gagaggacca tcagtttcaa ggacgacggc
     1561 acctacaaga cacgggctga ggtgaaattc gagggtgata cactcgtgaa tcgtatcgag
     1621 ctgaagggca tagattttaa ggaggatggc aatatcctgg gccacaagtt ggagtataac
     1681 ttcaacagtc acaatgtcta tataactgcc gataagcaga agaatggcat taaagccaat
     1741 ttcaaaattc gtcacaatgt cgaaggcggc gcagaacaga agctcatcag tgaagaggat
     1801 ctggccgcag agcagaaact cattagcgag gaggacttgg gcggaggcgg cgagcagaag
     1861 ctgatatcgg aggaagacct ggcggccgag caaaagctca tctcggagga ggatctcgcg
     1921 ggaggagatg gaagtgtcca gttggcggat cactaccagc agaatacgcc aattggtgac
     1981 ggccccgtcc ttctgccgga taaccactac ctctccaccc agtcggtgct gagcaaggac
     2041 ccgaatgaaa aacgtgacca catggttctt cttgagttcg tcacggctgc gggcatcacg
     2101 ttgggcatgg acgaactgta caagggcgct gaacagaaac ttatctccga ggaggatctg
     2161 gcggaacaga agctgatcag cgaagaggac ctggcggagc agaaacttat cagcgaagag
     2221 gatctgtaaa ccggagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     2281 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2341 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2401 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2461 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2521 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     2581 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     2641 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     2701 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     2761 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2821 ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag
     2881 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2941 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3001 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     3061 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     3121 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     3181 tggccactcc ctctctgcgc gctcgctcgc tcactgacgg ccgggcgmcc mwmggtcssc
     3241 csgcsycmgy gmrckgrsgc cgggcgacca aaggtcgccc ggcctcagtg agcgagcgag
     3301 cgcgcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3961 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4261 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4321 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4381 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4441 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4501 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4561 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4621 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4681 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4741 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4801 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4861 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4921 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4981 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5041 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5101 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5161 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5221 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5281 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5341 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5401 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5461 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5521 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5581 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5641 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5701 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     5761 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5821 ctgcgcgctc gctcgctcac tgaggccgcc cgggcrargc ccgggcgntc ggtgcggscc
     5881 tcttcgctat tacgcc
//
