LOCUS       sequence-189889-        6076 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cre dependent AAV expression of GFP spaghetti monster myc construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6076)
  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 6076)
  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: pENN.AAV.CAG.Flex.GFPsm_myc.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6076
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     395..417
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     479..498
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    533..566
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    629..662
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(676..705)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(709..738)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(742..771)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(985..1014)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1021..1050)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1063..1092)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1099..1128)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1660..1689)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1693..1722)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1726..1755)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1755..1764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1785..1818)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1889..1922)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1964..2552
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2017..2037)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2435..2446)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2595..2716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2606..2625)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2660..2679
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2878..3007
                     /label=AAV2 ITR
     rep_origin      complement(3244..3832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3324..3343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4003..4794,4795..4863))
                     /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     4626..4645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4864..4968)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4994..5449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5131..5152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5343..5362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5521..5650
                     /label=AAV2 ITR
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      301 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
      361 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
      421 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
      481 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcc ccataacttc
      541 gtataaagta tcctatacga agttatatca aaataggaag accaatgctt caccatcgac
      601 ccgaattgcc aagcatcacc atcgacccat aacttcgtat aatgtatgct atacgaagtt
      661 atactagccg gtttacagat cctcttcgct gataagtttc tgctccgcca ggtcctcttc
      721 gctgatcagc ttctgttccg ccagatcctc ctcggagata agtttctgtt cagcgccctt
      781 gtacagttcg tccatgccca acgtgatgcc cgcagccgtg acgaactcaa gaagaaccat
      841 gtggtcacgt ttttcattcg ggtccttgct cagcaccgac tgggtggaga ggtagtggtt
      901 atccggcaga aggacggggc cgtcaccaat tggcgtattc tgctggtagt gatccgccaa
      961 ctggacactt ccatctcctc ccgcgagatc ctcctccgag atgagctttt gctcggccgc
     1021 caggtcttcc tccgatatca gcttctgctc gccgcctccg cccaagtcct cctcgctaat
     1081 gagtttctgc tctgcggcca gatcctcttc actgatgagc ttctgttctg cgccgccttc
     1141 gacattgtga cgaattttga aattggcttt aatgccattc ttctgcttat cggcagttat
     1201 atagacattg tgactgttga agttatactc caacttgtgg cccaggatat tgccatcctc
     1261 cttaaaatct atgcccttca gctcgatacg attcacgagt gtatcaccct cgaatttcac
     1321 ctcagcccgt gtcttgtagg tgccgtcgtc cttgaaactg atggtcctct cttgaacgta
     1381 accctcgggc atagcgctct tgaaaaagtc atgctgcttc atgtgatctg gataccgcga
     1441 aaaacactga actccgccac ccaaggttgt gaccagggtg ggccagggta cgggcagttt
     1501 acccgtggtg caaatgaact tcagggtcag tttaccgttg gttgcgtcgc cctcgccctc
     1561 tccccgtacg gagaatttat gtccattgac atctccgtcc agctcgacga ggatgggcac
     1621 cacgccggtg aacagctctt cacccttcga cactccggca aggtcttcct cactgatcag
     1681 tttttgttcg gcaagatcct cttccgaaat cagtttctgc tctgccaagt cttcctccga
     1741 tatcagtttt tgctccatgg tggcggccgc aattgcgcta gagtataact tcgtatagga
     1801 tactttatac gaagttatca ttgggattct tcctattttg atccaagcat caccatcgac
     1861 cctctagtcc agatctcacc atcgacccat aacttcgtat agcatacatt atacgaagtt
     1921 atgtccctcg aagaggttcg aattcgatat caagcttatc gataatcaac ctctggatta
     1981 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2041 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2101 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2161 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2221 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2281 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2341 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     2401 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2461 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2521 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct
     2581 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2641 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2701 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2761 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     2821 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     2881 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2941 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3001 cgcgcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4981 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5041 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5101 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5161 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5221 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5281 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5341 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5401 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     5461 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5521 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
     5581 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     5641 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
     5701 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5761 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5821 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     5881 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     5941 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     6001 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     6061 gggggcgggg gggggg
//
