LOCUS       sequence-423178-        5722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  double floxed, myristoylation site (myr) and LDLR, fused to 
            reversibly switchable EGFP rsEGFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5722)
  AUTHORS   Willig KI, Wegner W, Muller A, Calvet-Fournier V, Steffens H
  TITLE     Multi-label in vivo STED microscopy by parallelized switching of 
            reversibly switchable fluorescent proteins.
  JOURNAL   Cell Rep. 2021 Jun 1;35(9):109192. doi: 
            10.1016/j.celrep.2021.109192.
  PUBMED    34077731
REFERENCE   2  (bases 1 to 5722)
  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: "Cell Rep.";
            date: "2021-06-1"; volume: "35(9):109192. doi"; pages: " 
            10.1016/j.celrep.2021.109192"
COMMENT     Sequence Label: pAAV-hSyn-DIO-myr-rsEGFP2- LDLR
FEATURES             Location/Qualifiers
     source          1..5722
                     /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        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(654..687)
                     /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(738..771)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(939..1655)
                     /codon_start=1
                     /product="reversibly and rapidly switchable enhanced green 
                     fluorescent protein (Grotjohann et al., 2012)"
                     /label=rsEGFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLAYGVLCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     SNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(981..1002)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1330..1349
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1589..1610
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    1707..1740
                     /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    1791..1824
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1849..2437
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1902..1922)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2320..2331)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2469..2945
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2668..2687)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2985..3125
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2985..3114
                     /label=AAV2 ITR
     rep_origin      3200..3655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3287..3306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3497..3518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3672..3691)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3791..3813
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3851..3869)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3937..4041
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4042..4110,4111..4902)
                     /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(4260..4279)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5073..5661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5562..5581
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc cgtcgactcc ggaataactt
      661 cgtataggat actttatacg aagttatgca gaatggtagc tggattgtag ctgctattag
      721 caatatgaaa cctcttaata acttcgtata gcatacatta tacgaagtta tggcgcgcct
      781 caggccacat cgtcctcgag gctgaccatc tgccgacttg gataggtgta cccgtcttgg
      841 gacctgcaaa tgtgcagctc atcctctgtg gtcttctggt acacggggtt atcgaaattg
      901 atgctgttga tattcttcag gcgccagttc cgaccggtct tgtacagctc gtccatgccg
      961 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     1021 gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     1081 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     1141 atgttgtggc ggatcttgaa gttagacttg atgccgttct tctgcttgtc ggccatgata
     1201 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     1261 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     1321 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     1381 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     1441 aagcacagca cgccgtaggc cagggtggtc acgagggtgg gccagggcac gggcagcttg
     1501 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     1561 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     1621 accccggtga acagctcctc gcccttgctc accatcgtca gttttgttgc ttctttatcc
     1681 ttacattgca cacagcccat gctagcataa cttcgtataa agtatcctat acgaagttat
     1741 ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt
     1801 ataatgtatg ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg
     1861 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1921 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1981 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2041 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2101 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2161 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2221 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     2281 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2341 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2401 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     2461 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     2521 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     2581 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     2641 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     2701 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2761 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2821 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2881 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     2941 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     3001 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     3061 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3121 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3181 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3241 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     3301 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3361 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     3421 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3481 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     3541 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     3601 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     3661 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     3721 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3781 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3841 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3901 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3961 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4021 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4081 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4141 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4201 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4261 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4321 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4381 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4441 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4501 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4561 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4621 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4681 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4741 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4801 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4861 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4921 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4981 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5041 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5101 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5161 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5221 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5281 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5341 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5401 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5461 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5521 ggcagggtcg gaacaggaga gcgcatgagg gagcttccag ggggaaacgc ctggtatctt
     5581 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5641 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5701 tgctggcctt ttgctcacat gt
//
