LOCUS       sequence-455990-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adeno-associated virus plasmid, Cre-dependent FLEX switch, APEX2NES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  AUTHORS   Joesch M, Mankus D, Yamagata M, Shahbazi A, Shalek R, Suissa-Peleg 
            A, Meister M, Lichtman JW, Scheirer WJ, Sanes JR
  TITLE     Reconstruction of genetically identified neurons imaged by 
            serial-section electron microscopy.
  JOURNAL   Elife. 2016 Jul 7;5. pii: e15015. doi: 10.7554/eLife.15015.
  PUBMED    27383271
REFERENCE   2  (bases 1 to 6890)
  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: "Elife."; 
            date: "2016-07-7"; volume: "5. pii: e15015. doi"; pages: " 
            10.7554/eLife.15015"
COMMENT     Sequence Label: AAV-CAG-DIO-APEX2NES
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        795..1174
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1454..2470
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2394..2416
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2478..2497
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2549..2582)
                     /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(2633..2666)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2681..2707)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(3467..3508)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      3508..3517
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3525..3558
                     /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    3609..3642
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3667..4255
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3720..3740)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4138..4149)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4287..4763
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4486..4505)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4803..4943
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4803..4932
                     /label=AAV2 ITR
     rep_origin      5018..5473
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5105..5124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5315..5336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5490..5509)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5609..5631
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5669..5687)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5755..5859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5860..5928,5929..6720)
                     /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(6078..6097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gctcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      841 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      901 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      961 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1021 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1081 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1141 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
     1201 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
     1261 attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccggggg gggggggggg
     1321 gggggggggg gggggggggg sgrggsggrg aggkgcggcg gcagccaatc agagcggcgc
     1381 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     1441 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
     1501 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     1561 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
     1621 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
     1681 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
     1741 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
     1801 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
     1861 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     1921 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     1981 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     2041 gccgggcggg gcggggccgc ckcgggccgg ggrgggctcg ggggrggggc gcggcggccc
     2101 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     2161 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     2221 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     2281 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     2341 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     2401 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     2461 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     2521 attcggatcc tctagagtcg actccggaat aacttcgtat aggatacttt atacgaagtt
     2581 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc
     2641 gtatagcata cattatacga agttatggcg cgccctagtc cagggtcagg cgctccaggg
     2701 gtggcagctg cagggcgtcg gcaaatccca gttctgacag cttctgatgt gcttctgcgt
     2761 aatctgcgaa aaaagcatcc tcgtcggctg catacttatc gaccagaggt ctgaacacgg
     2821 gatcggacag cagggctttg tcgctgggca gctgcagcag cccttccttc tctccactca
     2881 gcagttcagt aaagtaagaa ttgtcgaaaa tcagggggtt gcttgtccag ggtccctcga
     2941 agcctgacct ttccttatga gcggcgccaa tggtgtgccc tccagacagt gcgacgatat
     3001 cctggtcagt caggcccata gctttcccaa acacatctcg cagatggtcg gagccctttg
     3061 tcggatctgg caggcgcccc tctggaggtg gttctggttt gtcctcccgt cctgggtgga
     3121 aaggcacctt ggggccccca gtgacctcca cagcgaccac tcctgccagc tgataaaaat
     3181 cggcgtagga cagaattggg aactctgctt tcagaggttc cagcagccgc acggcgatgt
     3241 ccagcccatt gttggcacta tgtgccagtt cagcagggtg cttgattgtt ccaaagggtc
     3301 cgcctgtttt ggttcccttg tcaaaggtgc ctgcgctgtg gaaagccagt ctcagcatca
     3361 gaggtgcgca cctcttttct gcgataaacc ctctcagctt cttcttggct ttctccacgg
     3421 catcctggta gtctgcggac actgttgggt atgatttccc accggtggtg ctgtccaggc
     3481 ccagcagggg gttggggatg ggcttgccca tggtggcggc tagcataact tcgtataaag
     3541 tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg
     3601 tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga tatcaagctt
     3661 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3721 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3781 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3841 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3901 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3961 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4021 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4081 ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4141 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4201 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     4261 taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc
     4321 tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt
     4381 gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat
     4441 ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa
     4501 gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga
     4561 ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct
     4621 aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact
     4681 cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa
     4741 ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc
     4801 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     4861 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     4921 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     4981 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     5041 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     5101 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5161 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5221 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5281 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5341 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     5401 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5461 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5521 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5581 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5641 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5701 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5821 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5881 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5941 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6001 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6061 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6121 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6181 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6241 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6301 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6361 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6421 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6481 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6541 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6601 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6661 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6721 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6781 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6841 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
