LOCUS       sequence-348372-        3109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low Copy Number DNA Donor Vector for Bxb1 Integrase-mediated 
            Cassette Exchange (for inserts >15kb) into dual site mouse strains 
            (Bxb-GT/GA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3109)
  AUTHORS   Low BE, Hosur V, Lesbirel S, Wiles MV
  TITLE     Efficient targeted transgenesis of large donor DNA into multiple 
            mouse genetic backgrounds using bacteriophage Bxb1 integrase.
  JOURNAL   Sci Rep. 2022 Mar 31;12(1):5424. doi: 10.1038/s41598-022-09445-w.
  PUBMED    35361849
REFERENCE   2  (bases 1 to 3109)
  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: "Sci Rep."; 
            date: "2022-03-31"; volume: "12(1):5424. doi"; pages: " 
            10.1038/s41598-022-09445-w"
COMMENT     Sequence Label: p5155
FEATURES             Location/Qualifiers
     source          1..3109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..30)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    116..256
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     171..190
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(271..288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(442..1030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(522..541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1207..2001)
                     /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(1318..1337)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1928..1947
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2002..2106)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    2129..2166
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    2246..2283
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             join(2932..3109,1..14)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
       61 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
      121 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
      181 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      241 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      301 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      361 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      421 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      481 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      541 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      601 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      661 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      721 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      781 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      841 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
      901 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1021 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1081 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1141 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1201 aagctttcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     1261 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     1321 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     1381 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     1441 ggtcacgacg agatcctcgc cgtcgggcat ccgcgccttg agcctggcga acagttcggc
     1501 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     1561 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     1621 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     1681 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     1741 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     1801 tagccgcgct gcctcgtctt ggagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     1861 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     1921 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     1981 caatccatct tgttcaatca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2041 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2101 tccgcgcaca tttccccgac cggtggccgg cttgtcgacg acggcggtct ccgtcgtcag
     2161 gatcatccgg atccagtact ggtaccgcta gcagatatcg agctcgcggc cgcctcgagt
     2221 ctagagttta aacttaatta aggccggctt gtcgacgacg gcggactccg tcgtcaggat
     2281 catccgggaa ttcatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt
     2341 ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata
     2401 tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc
     2461 tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
     2521 tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc
     2581 tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa
     2641 agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga
     2701 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc
     2761 tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt
     2821 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc
     2881 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa
     2941 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg
     3001 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac
     3061 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacg
//
