LOCUS       sequence-111447-        5319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Bxb1-mediated recombination of the GFP-Dnmt3b1 cDNA into
            a MIN-tagged locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5319)
  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 5319)
  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-Dnmt3b1-Poly(A)
FEATURES             Location/Qualifiers
     source          1..5319
                     /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    3434..3568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3458..3477)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3512..3531
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3704..3751
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3770..4573
                     /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(3833..3852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4443..4462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4702..5290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4782..4801)
                     /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 acaggcccgt gatggcttca aagaatgata agctcgagct gcaggactgc
     3121 ctggagttca gtaggacagc aaagttaaag aaagtgcaga caataaccac caagtcgaac
     3181 tccatcagac agggcaaaaa ccagcttttc cctgtagtca tgaatggcaa ggacgacgtt
     3241 ttgtggtgca ctgagctcga aaggatcttc ggcttccctg ctcactacac ggacgtgtcc
     3301 aacatgggcc gcggcgcccg tcagaagctg ctgggcaggt cctggagtgt accggtcatc
     3361 agacacctgt ttgccccctt gaaggactac tttgcctgtg aataggcggc cgcagttaac
     3421 gaattctcta gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     3481 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3541 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3601 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     3661 ggctgattat gatcctgcaa gcctcgtcgc cgcggtttat tctgttgaca attaatcatc
     3721 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tgggatcggc
     3781 cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3841 ctatgactgg gcacaacaga cgatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3901 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3961 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4021 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4081 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4141 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4201 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4261 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4321 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4381 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4441 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4501 cgtgctttac ggtatcgccg cccccgattc gcagcgcatc gccttctatc gccttcttga
     4561 cgagttcttc tgagcgggac tctggggttc gaataaagac cgaccaagcg acgtctgaga
     4621 gctccctggc gaattcggta ccaataaaag agctttattt tcatgatctg tgtgttggtt
     4681 tttggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5101 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5281 aggatctcaa gaagatcctt tgatcttttc tagtgtgcg
//
