LOCUS       sequence-328182-        2777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector backbone to support a knock-in of 
            Linker-mTurquoise2 at the C-terminus of a target locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2777)
  AUTHORS   Bollen Y, Hageman JH, van Leenen P, Derks LLM, Ponsioen B, Buissant 
            des Amorie JR, Verlaan-Klink I, van den Bos M, Terstappen LWMM, van 
            Boxtel R, Snippert HJG
  TITLE     Efficient and error-free fluorescent gene tagging in human organoids
            without double-strand DNA cleavage.
  JOURNAL   PLoS Biol. 2022 Jan 28;20(1):e3001527. doi: 
            10.1371/journal.pbio.3001527.
  PUBMED    35089911
REFERENCE   2  (bases 1 to 2777)
  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: "PLoS 
            Biol."; date: "2022-01-28"; volume: "20(1):e3001527. doi"; pages: " 
            10.1371/journal.pbio.3001527"
COMMENT     Sequence Label: TVBB C-term-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..2777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..64)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(218..806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(298..317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(977..1768,1769..1837))
                     /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     1600..1619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1838..1942)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2112..2133)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2373..2392)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2720..2741
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tacaagtgac gaagagcgtt taaacgcgaa ccggctcttc gaattcctcg ctcactgact
       61 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
      121 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      181 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      241 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      301 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      361 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      421 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      481 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      541 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      601 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      661 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      721 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      781 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      841 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      901 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      961 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1021 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1081 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1141 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1201 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1261 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1321 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1381 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1441 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1501 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1561 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1621 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1681 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1741 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1801 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1861 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1921 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa
     1981 ataccgcaca gatgcgtaag gagaaaatac cgctacggaa gagcgcgatc gcgtttaaag
     2041 gctcttcagg cggaggcggc agcgccagcg tgagcaaggg cgaggagctg ttcaccgggg
     2101 tggtgcccat cctggtcgag ctggacggcg acgtaaatgg ccacaagttc agcgtgtccg
     2161 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2221 gaaagctgcc tgtgccctgg ccaaccctcg tgaccaccct gtcttggggc gtgcagtgct
     2281 tcgccagata ccctgaccac atgaagcagc acgacttctt caagtctgcc atgcccgaag
     2341 gctacgtcca ggagaggacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2401 aggtgaagtt cgagggcgac accctggtga acagaatcga gctgaagggc atcgacttca
     2461 aggaggacgg aaacatcctg gggcacaagc tggagtacaa ctacttctct gacaacgtct
     2521 atatcaccgc cgacaagcag aagaatggca tcaaggccaa cttcaagatc cgccacaaca
     2581 tcgaggatgg cggagtgcag ctcgccgatc actaccagca gaatacccct atcggcgacg
     2641 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
     2701 ccaacgagaa gagagatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2761 tcggaatgga cgagctg
//
