LOCUS       sequence-235213-        3049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate donor vector for the assembly of EGFP-based mammalian 
            recombinase reporters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3049)
  AUTHORS   Bessen JL, Afeyan LK, Dancik V, Koblan LW, Thompson DB, Leichner C, 
            Clemons PA, Liu DR
  TITLE     High-resolution specificity profiling and off-target prediction for 
            site-specific DNA recombinases.
  JOURNAL   Nat Commun. 2019 Apr 26;10(1):1937. doi: 10.1038/s41467-019-09987-0.
  PUBMED    31028261
REFERENCE   2  (bases 1 to 3049)
  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: "Nat 
            Commun."; date: "2019-04-26"; volume: "10(1):1937. doi"; pages: " 
            10.1038/s41467-019-09987-0"
COMMENT     Sequence Label: pBT100-neo
FEATURES             Location/Qualifiers
     source          1..3049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     227..244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             296..1090
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(350..369)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     960..979
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1270..1404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1307..1326)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1361..1380
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1597..2412)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1597..2412)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2320..2339
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2404..2423
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2534..3049,1..73)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3023..3042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
       61 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      121 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      181 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      241 cgagaatgct cttggtctca tgcctgatca agagacagga tgaggatcgt ttcgcatgat
      301 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
      361 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
      421 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
      481 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
      541 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
      601 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
      661 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
      721 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
      781 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
      841 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
      901 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
      961 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     1021 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     1081 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     1141 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     1201 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1261 cccatcgata acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1321 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1381 aatgtatctt atcatgtctg gatcaaatcc gaacgcagca atgagaccaa gagcctggtt
     1441 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     1501 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1561 atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa
     1621 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1681 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1741 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1801 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1861 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1921 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagccagac gaaatacgcg
     1981 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2041 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2101 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2161 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2221 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2281 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2341 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     2401 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     2461 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2521 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2581 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3001 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcc
//
