LOCUS       sequence-122104-        2746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the generation of gene disruptions via Bxb1-mediated 
            recombination into MIN-tagged cell lines. This vector can also be 
            used to express cDNAs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2746)
  AUTHORS   Mulholland CB, Smets M, Schmidtmann E, Leidescher S, Markaki Y, 
            Hofweber M, Qin W, Manzo M, Kremmer E, Thanisch K, Bauer C, Rombaut 
            P, Herzog F, Leonhardt H, Bultmann S
  TITLE     A modular open platform for systematic functional studies under 
            physiological conditions.
  JOURNAL   Nucleic Acids Res. 2015 May 24. pii: gkv550.
  PUBMED    26007658
REFERENCE   2  (bases 1 to 2746)
  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: "Nucleic 
            Acids Res. 2015 May 24. pii: gkv550."
COMMENT     Sequence Label: attB-mCh-Poly(A)
FEATURES             Location/Qualifiers
     source          1..2746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    9..46
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             49..753
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(191..209)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     459..478
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      799..808
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    861..995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(885..904)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     939..958
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1131..1178
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1197..2000
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPGGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1260..1279)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1870..1889
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(2129..2717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2209..2228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatatcccgg cttgtcgacg acggcggtct ccgtcgtcag gatcatccgt gagcaagggc
       61 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
      121 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      181 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
      241 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
      301 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      361 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
      421 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
      481 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
      541 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
      601 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
      661 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
      721 cactccaccg gcggcatgga cgagctgtac aagagcggcc tgaggagcag agcccaggcg
      781 agcaacagcg ccgtggacgc caccatgggc gatcgcccgg gaattgacta gtgcggccgc
      841 agttaacgaa ttctctagag gatccagaca tgataagata cattgatgag tttggacaaa
      901 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      961 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1021 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1081 gtggtatggc tgattatgat cctgcaagcc tcgtcgccgc ggtttattct gttgacaatt
     1141 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     1201 gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     1261 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     1321 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     1381 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     1441 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     1501 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     1561 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     1621 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     1681 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     1741 ccggcggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     1801 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     1861 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     1921 gcttcctcgt gctttacggt atcgccgccc ccgattcgca gcgcatcgcc ttctatcgcc
     1981 ttcttgacga gttcttctga gcgggactct ggggttcgaa taaagaccga ccaagcgacg
     2041 tctgagagct ccctggcgaa ttcggtacca ataaaagagc tttattttca tgatctgtgt
     2101 gttggttttt ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2701 gaaaaaaagg atctcaagaa gatcctttga tcttttctag tgtgcg
//
