LOCUS       sequence-149903-        7104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adeno-associated virus plasmid, Cre-dependent FLEX switch, 
            erHRP(N175S mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  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 7104)
  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-erHRP(N175S mutant)
FEATURES             Location/Qualifiers
     source          1..7104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        156..535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        537..814
                     /label=chicken beta-actin promoter
     intron          816..1833
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1757..1779
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1841..1860
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1912..1945)
                     /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(1996..2029)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2986..3012)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(3016..3075)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3072..3081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3089..3122
                     /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    3173..3206
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3231..3819
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3284..3304)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3702..3713)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3851..4327
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4050..4069)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4367..4507
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4367..4496
                     /label=AAV2 ITR
     rep_origin      4582..5037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4669..4688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4879..4900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5054..5073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5173..5195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5233..5251)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5319..5423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5424..5492,5493..6284)
                     /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(5642..5661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6455..7043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6944..6963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgctcgacat tgattattga ctagttatta
      181 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      241 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      301 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      361 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      421 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      481 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
      541 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
      601 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
      661 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
      721 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
      781 gcggccctat aaaaagcgaa gcgcgcggcg ggcggggagt cgctgcgacg ctgccttcgc
      841 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      901 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      961 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
     1021 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
     1081 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     1141 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
     1201 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
     1261 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
     1321 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
     1381 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc
     1441 tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1501 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1561 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1621 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1681 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1741 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1801 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1861 ctgtctcatc attttggcaa agaattcgga tcctctagag tcgactccgg aataacttcg
     1921 tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct gctattagca
     1981 atatgaaacc tcttaataac ttcgtatagc atacattata cgaagttatg gcgcgcctta
     2041 aagttcgtcc ttcaagtcag agttgctgtt cacgacccta cagttcagtc ttatctgtcc
     2101 ctgcgtaccc gtcaggggcg ttatatttcc catccgatcc atagcttcca cgaaagcatt
     2161 gaaaaaggtt tgtgtgctat tagcgaagct gcgcacaaga ggaatagtgt cggtagcatt
     2221 tggggaactg aacagctcct ggtcggactg aatcaggccc ttctgttcct ccagattgac
     2281 gtaatactta ttgtcgaaga tggttggggt gcgcaggtca aagtcaacca gtgcagacaa
     2341 gttgccattg agcggacaca gccctctgag agtctgcaag tatgttgtgt tcagggtagg
     2401 gtcaggcaag ccggtgttgg aaaagttata cagtctgtcc atgatgaacc tgcactgact
     2461 cttgccgaat gtgtggcctc cggacagagc caccaaatca ctacttcggt tcaggccgac
     2521 attcctgaag gaatccttca gctggggaag ggtgaagaag ggagcaggca ggtttgcgtt
     2581 tgccaagtcc agaaaagcct gcagagagtc cctccttccc agggggaccc tccagctggg
     2641 gccaccggcc agcgtcacgg attgctgggc cgctatggtc agaaggtctg cacaactaac
     2701 tgtccggggg caggcgctct ccacagcggc tttcattcta tcgatcacgg gaaaccctct
     2761 agcactattc gcgttaccga aggcatcctt ttcagtcctg aaagaggtgg tattgtcgag
     2821 gaggatggaa gcatcacagc cgtttacaaa acaatcgtgg aaatgcagcc gcaaaattga
     2881 agcggctatc ctcggatcag accgcaactc gttcacgata gtatctcgca cgatgttgct
     2941 cacgttagga caagagttat cgtagaaagt tggagtaagc tgcatagcat aatctggaac
     3001 atcatatgga tagtcaccag tggaacctgg aacccagagc agcagtaccc atagcaggag
     3061 tgtgtctgtc tccatggtgg cggctagcat aacttcgtat aaagtatcct atacgaagtt
     3121 atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc
     3181 gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat aatcaacctc
     3241 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3301 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3361 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3421 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3481 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3541 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3601 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
     3661 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3721 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3781 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
     3841 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
     3901 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
     3961 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
     4021 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
     4081 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
     4141 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
     4201 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
     4261 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
     4321 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
     4381 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4441 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4501 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4561 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4621 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4681 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4741 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4801 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4861 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4921 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4981 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     5041 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5101 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5161 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5221 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5281 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5401 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5461 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5521 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5581 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5641 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5701 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5761 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5821 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5881 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5941 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6001 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6061 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6121 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6181 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6241 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6601 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7081 tttgctggcc ttttgctcac atgt
//
