LOCUS       sequence-360209-        4762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CreON FlpOFF knock-in for smFP-HA-CaV2.3 (amino acid position: G5).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4762)
  AUTHORS   Droogers WJ, Willems J, MacGillavry HD, de Jong APH
  TITLE     Duplex Labeling and Manipulation of Neuronal Proteins Using 
            Sequential CRISPR/Cas9 Gene Editing.
  JOURNAL   eNeuro. 2022 Jul 18;9(4):ENEURO.0056-22.2022. doi: 
            10.1523/ENEURO.0056-22.2022.
  PUBMED    35851300
REFERENCE   2  (bases 1 to 4762)
  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: 
            "10.1523/ENEURO.0056-22"; journalName: "eNeuro"; date: "2022-07-18- 
            18"; volume: "9"; issue: "4"; pages: "ENEURO.0056-22.2022"
COMMENT     Sequence Label: pOC3 smFP-HA-Cacna1e KI
FEATURES             Location/Qualifiers
     source          1..4762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    39..72
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(343..376)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     protein_bind    complement(607..640)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     protein_bind    688..721
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             926..952
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             956..982
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             986..1012
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1541..1567
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1574..1600
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1613..1639
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1646..1672
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1883..1909
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1913..1939
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1943..1969
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   2067..2192
                     /label=AAV2 ITR
     rep_origin      2278..2733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2365..2384)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2575..2596
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2750..2769)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2869..2891
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2929..2947)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3015..3119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3120..3188,3189..3980)
                     /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(3338..3357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4151..4739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4640..4659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatgtga agttcctatt ctctagaaag
       61 tataggaact tcgatccgac gccgccatct ctaggcccgc gccggccccc tcgcacagac
      121 ttgtgggaga agctcggcta ctcccctgcc ccggttaatt tgcatataat atttcctagt
      181 aactatagag gcttaatgtg cgataaaaga cagataatct gttcttttta atactagcta
      241 cattttacat gataggcttg gatttctata agagatacaa atactaaatt attattttaa
      301 aaaacagcac actcgagaac tcaccctaac tgtaaagtta gtataacttc gtatagtata
      361 aattatacga agttataagc cttgtttttt gcgatttttg aattcgttcc tcagaggaac
      421 tgacaagcac cctaacatcc tattggaggc tcactcacgt tttttctatt ttgtttcttg
      481 acagcagagc tcgttgctca ctgtatagct caggttggcc tgacactgat gaggttctcc
      541 agtgactgcc tctacctacc tactgggatg acagaggtgt accaccaagc cacgcccggg
      601 ggatccataa cttcgtatag tataaattat acgaagttat aagccttgtg caggatggct
      661 cgcttcgggg gttttagagc tatgctggaa gttcctattc tctagaaagt ataggaactt
      721 ccagcatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      781 cggtgctttt ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta
      841 gcgcgtgcgc caattctgca gacaaatggc tctagaagct tcccccccga agcgagccat
      901 cctgatggct agcggagcca ccatgtaccc ttatgatgtg cccgattatg ctggctaccc
      961 ttatgatgtt cctgattacg ccggatatcc gtatgacgtg ccagactacg cgggaggtgt
     1021 gagcaaggga gaggagttgt ttaccggcgt ggtccccata ctggtggagt tggatggcga
     1081 cgttaatgga cataaattct cggtccgcgg cgagggagag ggagacgcca ccaatggcaa
     1141 gctgacgctt aagttcattt gcacgactgg aaaattgcct gtcccctggc cgaccctggt
     1201 caccacactg ggcggaggag tgcagtgctt ctcccgttac ccagaccaca tgaagcagca
     1261 cgatttcttc aagagcgcaa tgccggaggg atacgtgcaa gaacggacta tttccttcaa
     1321 ggatgacgga acctataaga cccgtgcgga ggtcaaattc gagggtgaca ccctggtgaa
     1381 ccgaattgaa ctcaaaggaa tcgatttcaa ggaggatgga aatattctgg gtcacaagct
     1441 ggaatacaac ttcaacagcc ataatgtgta cattacggct gataagcaga agaacggcat
     1501 taaggccaat ttcaagatcc gccacaacgt tgagggtgga tatccctacg acgtgcccga
     1561 ttatgcgggc ggatatccgt atgatgttcc agattatgct ggtggtggag gctatcccta
     1621 tgatgtcccc gactacgccg gaggttaccc atacgacgtg cccgactacg ctggaggcga
     1681 tggcagcgtg cagctggcag atcattatca acagaatacc cccataggtg atggccccgt
     1741 tctgcttcca gataatcact acctttccac ccagagcgtg ctttcgaaag acccgaacga
     1801 aaagcgtgat cacatggtcc tgctggagtt tgtgaccgcg gctggaatca ccctgggtat
     1861 ggacgaactc tacaagggtg gctaccccta cgatgtgccg gattacgctg gatatccgta
     1921 tgacgtaccg gactatgccg gttacccgta cgatgtcccg gactacgctt caaccggtag
     1981 cgctgtttcc cccccgaagc gagccatcct gctcgactcg aggatcctag gacgcgttaa
     2041 cttacggtaa atggcccggc ggccgcagga acccctagtg atggagttgg ccactccctc
     2101 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     2161 tgcccgggcg gcctcagtga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     2221 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     2281 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2341 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2401 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2461 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2521 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2581 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     2641 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2701 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     2761 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2821 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2881 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2941 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3001 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3061 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3121 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3181 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3241 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3301 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3361 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3421 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3481 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3541 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3601 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3661 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3721 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3781 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc
     3841 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3901 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3961 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4021 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4741 acgccagcaa cgcggccttt tt
//
