LOCUS       sequence-274804-        3395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for PCR-generated donor DNA for rapid tagging of proteins 
            with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3395)
  AUTHORS   Paix A, Rasoloson D, Folkmann A, Seydoux G
  TITLE     Rapid Tagging of Human Proteins with Fluorescent Reporters by Genome
            Engineering using Double-Stranded DNA Donors.
  JOURNAL   Curr Protoc Mol Biol. 2019 Dec;129(1):e102. doi: 10.1002/cpmb.102.
  PUBMED    31710422
REFERENCE   2  (bases 1 to 3395)
  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: "Curr Protoc
            Mol Biol."; date: "2019-12"; volume: "129(1):e102. doi"; pages: " 
            10.1002/cpmb.102"
COMMENT     Sequence Label: pAP1837
FEATURES             Location/Qualifiers
     source          1..3395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     158..180
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     172..189
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     173..189
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(968..984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(968..984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(981..1003)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    992..1008
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1016..1022,1023..1040,1041..1046))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1061..1082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1199..1216)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1370..1958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1450..1469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2129..2920,2921..2989))
                     /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     2752..2771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2990..3094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3162..3180
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3218..3240)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3340..3359
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
       61 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      121 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      181 acggccagtg aattcgagct cggtacccgg ggatcgtctc taaaggagaa gaagataata
      241 tggctatcat taaagagttt atgcgattca aagtgcacat ggaaggaagt gtcaacggcc
      301 atgaattcga aatcgaaggg gaaggagaag gtcgccccta tgaaggcact cagacggcaa
      361 aacttaaggt tacaaaggga ggtccgcttc ccttcgcgtg ggatatactt tctcctcagt
      421 tcatgtatgg gtccaaagcc tacgtcaagc atccagcaga cattccggat tatctgaagc
      481 tgtcatttcc cgagggtttt aaatgggaaa gggtgatgaa tttcgaggac gggggagttg
      541 tgaccgttac acaggattct tcccttcagg acggcgagtt catatataaa gtcaagctcc
      601 gagggacaaa cttcccaagt gatggacctg tcatgcaaaa gaagaccatg ggctgggaag
      661 cttcttcaga acgaatgtat ccagaagacg gagcgctgaa aggtgagata aagcagaggt
      721 tgaaactgaa ggatgggggg cattacgatg cagaagtgaa aaccacatat aaagcaaaga
      781 aacccgtaca actgccagga gcatataatg tcaatatcaa acttgatatt accagccaca
      841 atgaagacta cactatagtt gagcaatacg aaagggcaga gggacggcat tccacaggcg
      901 gaatggatga actctataaa gatcctctag agtcgacctg caggcatgca agcttggcgt
      961 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1021 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1081 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1141 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1201 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1261 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1561 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2041 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2101 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2161 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2221 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2281 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2341 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2401 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2461 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2521 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2581 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2641 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2701 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2761 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2821 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2881 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2941 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3001 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3061 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3121 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     3181 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3241 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3301 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     3361 tgagagtgca ccatatgcgg tgtgaaatac cgcac
//
