LOCUS       sequence-238585-        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     75..92
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             144..938
                     /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(198..217)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     808..827
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1118..1252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1155..1174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1209..1228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1445..2260)
                     /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(1445..2260)
                     /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     2168..2187
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2252..2271
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2382..2970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2871..2890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
       61 gaacgaccga gcgcagcgag tcagtgagcg agaatgctct tggtctcatg cctgatcaag
      121 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
      181 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
      241 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
      301 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
      361 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
      421 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
      481 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
      541 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
      601 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
      661 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
      721 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
      781 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
      841 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
      901 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
      961 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     1021 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     1081 ggatctcatg ctggagttct tcgcccaccc catcgataac ttgtttattg cagcttataa
     1141 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1201 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaaatccga
     1261 acgcagcaat gagaccaaga gcctggttaa aaaatgagct gatttaacaa aaatttaacg
     1321 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     1381 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     1441 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1501 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1561 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1621 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1681 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     1741 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1801 attgcgcctg agccagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     1861 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     1921 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     1981 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2041 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2101 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     2161 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2221 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2281 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     2341 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2401 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2461 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2521 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2581 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2641 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2701 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2761 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2821 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2881 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2941 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3001 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatccc
//
