LOCUS       sequence-360144-        4484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CreON knock-in for Halo-Homer1 (amino acid position: G2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4484)
  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 4484)
  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: pOC2 CRISPIE-Halo-Homer1 KI
FEATURES             Location/Qualifiers
     source          1..4484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(39..72)
                     /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(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."
     misc_RNA        406..481
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             719..1609
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     repeat_region   1785..1925
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1785..1914
                     /label=AAV2 ITR
     rep_origin      2000..2455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2087..2106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2297..2318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2472..2491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2591..2613
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2651..2669)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2737..2841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2842..2910,2911..3702)
                     /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(3060..3079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3873..4461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4362..4381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatgtat aacttcgtat agtataaatt
       61 atacgaagtt atgatccgac 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 cttgttatat gcctccggtg cccaggtttt agagctagaa
      421 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      481 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
      541 tgcgccaatt ctgcagacaa atggctctag aagctttata tgcctccggt gcccaggggg
      601 agccagggaa taagacgtga aaggctgtcc tgccctggac actgggtttg gagctgagga
      661 ctgagcatgt gtgtctggtt ttgctttccc cctctgcagc aggagagcag ctagcggaat
      721 ggcagaaatc ggtactggct ttccattcga cccccattat gtggaagtcc tgggcgagcg
      781 catgcactac gtcgatgttg gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa
      841 cccgacctcc tcctacgtgt ggcgcaacat catcccgcat gttgcaccga cccatcgctg
      901 cattgctcca gacctgatcg gtatgggcaa atccgacaaa ccagacctgg gttatttctt
      961 cgacgaccac gtccgcttca tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt
     1021 cctggtcatt cacgactggg gctccgctct gggtttccac tgggccaagc gcaatccaga
     1081 gcgcgtcaaa ggtattgcat ttatggagtt catccgccct atcccgacct gggacgaatg
     1141 gccagaattt gcccgcgaga ccttccaggc cttccgcacc accgacgtcg gccgcaagct
     1201 gatcatcgat cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct
     1261 gactgaagtc gagatggacc attaccgcga gccgttcctg aatcctgttg accgcgagcc
     1321 actgtggcgc ttcccaaacg agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct
     1381 ggtcgaagaa tacatggact ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg
     1441 caccccaggc gttctgatcc caccggccga agccgctcgc ctggccaaaa gcctgcctaa
     1501 ctgcaaggct gtggacatcg gcccgggtct gaatctgctg caagaagaca acccggacct
     1561 gatcggcagc gagatcgcgc gctggctgtc cacgctcgag atttccggca gcgctgatca
     1621 tcgaaggtaa tccagtgcgt ctcccaggag gagtggccgt ccagacagcg gctattaaat
     1681 accaagactt caaagtgtca ggagctagga tgctatgata ttattctata tgcctccggt
     1741 gcccagggga cgcgttaact tacggtaaat ggcccggcgg ccgcaggaac ccctagtgat
     1801 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     1861 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     1921 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     1981 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2041 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     2101 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2161 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     2221 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2281 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     2341 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     2401 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     2521 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     2581 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     2641 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2701 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2821 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2881 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2941 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3001 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3061 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3121 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3181 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3241 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3301 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3361 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3421 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3481 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3541 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     3601 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3661 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3721 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3781 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3841 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3961 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4021 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4081 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4141 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4201 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4261 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4321 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4381 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4441 ggggggcgga gcctatggaa aaacgccagc aacgcggcct tttt
//
