LOCUS       sequence-111446-        5130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Bxb1-mediated recombination of the GFP-Dnmt3b6 cDNA into
            a MIN-tagged locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5130)
  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 5130)
  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-GFP-Dnmt3b6-Poly(A)
FEATURES             Location/Qualifiers
     source          1..5130
                     /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             join(49..51,52..762)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(49..51,52..762)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(91..112)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(352..371)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     699..720
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      808..817
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             897..908
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    3245..3379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3269..3288)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3323..3342
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3515..3562
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3581..4384
                     /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
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3644..3663)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4254..4273
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4513..5101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4593..4612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatatcccgg cttgtcgacg acggcggtct ccgtcgtcag gatcatccgt gagcaagggc
       61 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      121 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      181 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      241 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      301 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      361 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      421 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      481 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      541 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      601 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      661 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      721 accgccgccg ggatcactct cggcatggac gagctgtaca agagcggcct gaggagcaga
      781 gcccaggcga gcaacagcgc cgtggacgcc accatgggcg atcgcatgaa gggagacagc
      841 agacatctga atgaagaaga gggtgccagc gggtatgagg agtgcattat cgttaatggg
      901 aacttcagtg accagtcctc agacacgaag gatgctccct cacccccagt cttggaggca
      961 atctgcacag agccagtctg cacaccagag accagaggcc gcaggtcaag ctcccggctg
     1021 tctaagaggg aggtctccag ccttctgaat tacacgcagg acatgacagg agatggagac
     1081 agagatgatg aagtagatga tgggaatggc tctgatattc taatgccaaa gctcacccgt
     1141 gagaccaagg acaccaggac gcgctctgaa agcccggctg tccgaacccg acatagcaat
     1201 gggacctcca gcttggagag gcaaagagcc tcccccagaa tcacccgagg tcggcagggc
     1261 cgccaccatg tgcaggagta ccctgtggag tttccggcta ccaggtctcg gagacgtcga
     1321 gcatcgtctt cagcaagcac gccatggtca tcccctgcca gcgtcgactt catggaagaa
     1381 gtgacaccta agagcgtcag taccccatca gttgacttga gccaggatgg agatcaggag
     1441 ggtatggata ccacacaggt ggatgcagag agcagagatg gagacagcac agagtatcag
     1501 gatgataaag agtttggaat aggtgacctc gtgtggggaa agatcaaggg cttctcctgg
     1561 tggcctgcca tggtggtgtc ctggaaagcc acctccaagc gacaggccat gcccggaatg
     1621 cgctgggtac agtggtttgg tgatggcaag ttttctgaga tctctgctga caaactggtg
     1681 gctctggggc tgttcagcca gcactttaat ctggctacct tcaataagct ggtttcttat
     1741 aggaaggcca tgtaccacac tctggagaaa gccagggttc gagctggcaa gaccttctcc
     1801 agcagtcctg gagagtcact ggaggaccag ctgaagccca tgctggagtg ggcccacggt
     1861 ggcttcaagc ctactgggat cgagggcctc aaacccaaca agaagcaacc agtggttaat
     1921 aagtcgaagg tgcgtcgttc agacagtagg aacttagaac ccaggagacg cgagaacaaa
     1981 agtcgaagac gcacaaccaa tgactctgct gcttctgagt cccccccacc caagcgcctc
     2041 aagacaaata gctatggcgg gaaggaccga ggggaggatg aggagagccg agaacggatg
     2101 gcttctgaag tcaccaacaa caagggcaat ctggaagacc gctgtttgtc ctgtggaaag
     2161 aagaaccctg tgtccttcca ccccctcttt gagggtgggc tctgtcagag ttgccgggat
     2221 cgcttcctag agctcttcta catgtatgat gaggacggct atcagtccta ctgcaccgtg
     2281 tgctgtgagg gccgtgaact gctgctgtgc agtaacacaa gctgctgcag atgcttctgt
     2341 gtggagtgtc tggaggtgct ggtgggcgca ggcacagctg aggatgccaa gctgcaggaa
     2401 ccctggagct gctatatgtg cctccctcag cgctgccatg gggtcctccg acgcaggaaa
     2461 gattggaaca tgcgcctgca agacttcttc actactgatc ctgacctgga agaatttgag
     2521 ccacccaagt tgtacccagc aattcctgca gccaaaagga ggcccattag agtcctgtct
     2581 ctgtttgatg gaattgcaac ggggtacttg gtgctcaagg agttgggtat taaagtggaa
     2641 aagtacattg cctccgaagt ctgtgcagag tccatcgctg tgggaactgt taagcatgaa
     2701 ggccagatca aatatgtcaa tgacgtccgg aaaatcacca agaaaaatat tgaagagtgg
     2761 ggcccgttcg acttggtgat tggtggaagc ccatgcaatg atctctctaa cgtcaatcct
     2821 gcccgcaaag gtttatatga gggcacagga aggctcttct tcgagtttta ccacttgctg
     2881 aattataccc gccccaagga gggcgacaac cgtccattct tctggatgtt cgagaatgtt
     2941 gtggccatga aagtgaatga caagaaagac atctcaagat tcctggcatg taacccagtg
     3001 atgatcgatg ccatcaaggt gtctgctgct cacagggccc ggtacttctg gggtaaccta
     3061 cccggaatga acaggatctt cggcttccct gctcactaca cggacgtgtc caacatgggc
     3121 cgcggcgccc gtcagaagct gctgggcagg tcctggagtg taccggtcat cagacacctg
     3181 tttgccccct tgaaggacta ctttgcctgt gaataggcgg ccgcagttaa cgaattctct
     3241 agaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3301 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3361 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3421 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     3481 tgatcctgca agcctcgtcg ccgcggttta ttctgttgac aattaatcat cggcatagta
     3541 tatcggcata gtataatacg acaaggtgag gaactaaacc atgggatcgg ccattgaaca
     3601 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3661 ggcacaacag acgatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3721 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3781 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3841 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3901 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3961 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4021 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4081 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4141 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4201 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4261 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4321 cggtatcgcc gcccccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4381 ctgagcggga ctctggggtt cgaataaaga ccgaccaagc gacgtctgag agctccctgg
     4441 cgaattcggt accaataaaa gagctttatt ttcatgatct gtgtgttggt ttttggccgc
     4501 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4561 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4621 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4681 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4741 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4801 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4861 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4921 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4981 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5041 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5101 agaagatcct ttgatctttt ctagtgtgcg
//
