LOCUS       sequence-360125-        3649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CreON FlpOFF knock-in vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3649)
  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 3649)
  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 cloning template vector
FEATURES             Location/Qualifiers
     source          1..3649
                     /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    691..724
                     /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)"
     repeat_region   950..1090
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   950..1079
                     /label=AAV2 ITR
     rep_origin      1165..1620
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1252..1271)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1462..1483
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1637..1656)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1756..1778
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1816..1834)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1902..2006
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2007..2075,2076..2867)
                     /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(2225..2244)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3038..3626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3527..3546
                     /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 aagccttgtt tgtcttcgtg
      661 tacaagaaga cctgttttag agctatgctg gaagttccta ttctctagaa agtataggaa
      721 cttccagcat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
      781 agtcggtgct tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt
      841 ttagcgcgtg cgccaattct gcagacaaat ggctctagaa gcttgctagc aattgtcgac
      901 tcgaggatcc taggacgcgt taacttacgg taaatggccc ggcggccgca ggaaccccta
      961 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     1021 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     1081 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     1141 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1201 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     1261 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1321 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1381 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1441 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     1501 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     1561 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     1621 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     1681 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     1741 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     1801 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1861 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1921 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1981 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2041 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2101 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2161 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2221 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2281 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2341 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2401 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2461 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2521 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2581 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2641 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2701 aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta
     2761 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2821 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2881 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2941 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3001 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3061 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3121 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3181 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3241 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3301 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3361 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3421 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3481 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3541 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3601 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttt
//
