LOCUS       sequence-360225-        3693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CreON knock-in for GluA1-HA (amino acid position: stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3693)
  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 3693)
  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: pOC1 Gria1-HA KI
FEATURES             Location/Qualifiers
     source          1..3693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(309..342)
                     /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(573..606)
                     /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."
     misc_RNA        636..711
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             843..869
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             882..908
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   994..1134
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   994..1123
                     /label=AAV2 ITR
     rep_origin      1209..1664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1296..1315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1506..1527
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1681..1700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1800..1822
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1860..1878)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1946..2050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2051..2119,2120..2911)
                     /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(2269..2288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3082..3670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3571..3590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatgtga tccgacgccg ccatctctag
       61 gcccgcgccg gccccctcgc acagacttgt gggagaagct cggctactcc cctgccccgg
      121 ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga
      181 taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt tctataagag
      241 atacaaatac taaattatta ttttaaaaaa cagcacactc gagaactcac cctaactgta
      301 aagttagtat aacttcgtat agtataaatt atacgaagtt ataagccttg ttttttgcga
      361 tttttgaatt cgttcctcag aggaactgac aagcacccta acatcctatt ggaggctcac
      421 tcacgttttt tctattttgt ttcttgacag cagagctcgt tgctcactgt atagctcagg
      481 ttggcctgac actgatgagg ttctccagtg actgcctcta cctacctact gggatgacag
      541 aggtgtacca ccaagccacg cccgggggat ccataacttc gtatagtata aattatacga
      601 agttataagc cttgtgggag ccacaggatt gtaacgtttt agagctagaa atagcaagtt
      661 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
      721 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
      781 ctgcagacaa atggctctag aagcttaccc gttacaatcc tgtggctccc ggcgctagcg
      841 gttacccata cgatgttcca gattacgctg gatctggttc gtatccttat gacgtgcctg
      901 actacgcagg tggaagcgct cccgttacaa tcctgtggct cccacgcgtg gatcctagga
      961 ccggtcgggc catttaccgt aagttgcggc cgcaggaacc cctagtgatg gagttggcca
     1021 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     1081 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     1141 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     1201 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1261 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     1321 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1381 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     1441 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1501 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     1561 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     1621 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     1681 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     1741 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1801 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     1861 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1921 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1981 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2041 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2101 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2161 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2221 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2281 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2341 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2401 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2461 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2521 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2581 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2641 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2701 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2761 gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2821 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2881 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2941 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3001 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3061 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3661 cctatggaaa aacgccagca acgcggcctt ttt
//
