LOCUS       sequence-111441-        7124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for Bxb1-mediated recombination of the mCh-Dnmt3b1 cDNA into
            a MIN-tagged locus using Neomycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7124)
  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 7124)
  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-Dnmt3b1-Poly(A)-NeoR
FEATURES             Location/Qualifiers
     source          1..7124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..561
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(97..120)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     494..513
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             579..1373
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             579..1373
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1561..1580)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1605..1652
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    1823..1860
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attB site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     CDS             1863..2567
                     /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(2005..2023)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2273..2292
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      2613..2622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2702..2713
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    5239..5373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5263..5282)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5317..5336
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5509..5556
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5575..6378
                     /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"
     rep_origin      complement(6507..7095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6587..6606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatgccagct cattcctccc actcatgatc tatagatccc ccgggctgca ggaattctac
       61 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
      121 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
      181 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
      241 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
      301 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
      361 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
      421 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
      481 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
      541 tcctcatctc cgggcctttc gaccgatcat caagcttaat gattgaacaa gatggattgc
      601 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      661 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      721 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
      781 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      841 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      901 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      961 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     1021 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     1081 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     1141 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1201 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     1261 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1321 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     1381 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1441 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1501 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     1561 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     1621 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     1681 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     1741 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     1801 ggcaagctta catcgagatc ccggcttgtc gacgacggcg gtctccgtcg tcaggatcat
     1861 ccgtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1921 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1981 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     2041 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     2101 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     2161 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     2221 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     2281 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     2341 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     2401 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     2461 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     2521 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagagc ggcctgagga
     2581 gcagagccca ggcgagcaac agcgccgtgg acgccaccat gggcgatcgc atgaagggag
     2641 acagcagaca tctgaatgaa gaagagggtg ccagcgggta tgaggagtgc attatcgtta
     2701 atgggaactt cagtgaccag tcctcagaca cgaaggatgc tccctcaccc ccagtcttgg
     2761 aggcaatctg cacagagcca gtctgcacac cagagaccag aggccgcagg tcaagctccc
     2821 ggctgtctaa gagggaggtc tccagccttc tgaattacac gcaggacatg acaggagatg
     2881 gagacagaga tgatgaagta gatgatggga atggctctga tattctaatg ccaaagctca
     2941 cccgtgagac caaggacacc aggacgcgct ctgaaagccc ggctgtccga acccgacata
     3001 gcaatgggac ctccagcttg gagaggcaaa gagcctcccc cagaatcacc cgaggtcggc
     3061 agggccgcca ccatgtgcag gagtaccctg tggagtttcc ggctaccagg tctcggagac
     3121 gtcgagcatc gtcttcagca agcacgccat ggtcatcccc tgccagcgtc gacttcatgg
     3181 aagaagtgac acctaagagc gtcagtaccc catcagttga cttgagccag gatggagatc
     3241 aggagggtat ggataccaca caggtggatg cagagagcag agatggagac agcacagagt
     3301 atcaggatga taaagagttt ggaataggtg acctcgtgtg gggaaagatc aagggcttct
     3361 cctggtggcc tgccatggtg gtgtcctgga aagccacctc caagcgacag gccatgcccg
     3421 gaatgcgctg ggtacagtgg tttggtgatg gcaagttttc tgagatctct gctgacaaac
     3481 tggtggctct ggggctgttc agccagcact ttaatctggc taccttcaat aagctggttt
     3541 cttataggaa ggccatgtac cacactctgg agaaagccag ggttcgagct ggcaagacct
     3601 tctccagcag tcctggagag tcactggagg accagctgaa gcccatgctg gagtgggccc
     3661 acggtggctt caagcctact gggatcgagg gcctcaaacc caacaagaag caaccagtgg
     3721 ttaataagtc gaaggtgcgt cgttcagaca gtaggaactt agaacccagg agacgcgaga
     3781 acaaaagtcg aagacgcaca accaatgact ctgctgcttc tgagtccccc ccacccaagc
     3841 gcctcaagac aaatagctat ggcgggaagg accgagggga ggatgaggag agccgagaac
     3901 ggatggcttc tgaagtcacc aacaacaagg gcaatctgga agaccgctgt ttgtcctgtg
     3961 gaaagaagaa ccctgtgtcc ttccaccccc tctttgaggg tgggctctgt cagagttgcc
     4021 gggatcgctt cctagagctc ttctacatgt atgatgagga cggctatcag tcctactgca
     4081 ccgtgtgctg tgagggccgt gaactgctgc tgtgcagtaa cacaagctgc tgcagatgct
     4141 tctgtgtgga gtgtctggag gtgctggtgg gcgcaggcac agctgaggat gccaagctgc
     4201 aggaaccctg gagctgctat atgtgcctcc ctcagcgctg ccatggggtc ctccgacgca
     4261 ggaaagattg gaacatgcgc ctgcaagact tcttcactac tgatcctgac ctggaagaat
     4321 ttgagccacc caagttgtac ccagcaattc ctgcagccaa aaggaggccc attagagtcc
     4381 tgtctctgtt tgatggaatt gcaacggggt acttggtgct caaggagttg ggtattaaag
     4441 tggaaaagta cattgcctcc gaagtctgtg cagagtccat cgctgtggga actgttaagc
     4501 atgaaggcca gatcaaatat gtcaatgacg tccggaaaat caccaagaaa aatattgaag
     4561 agtggggccc gttcgacttg gtgattggtg gaagcccatg caatgatctc tctaacgtca
     4621 atcctgcccg caaaggttta tatgagggca caggaaggct cttcttcgag ttttaccact
     4681 tgctgaatta tacccgcccc aaggagggcg acaaccgtcc attcttctgg atgttcgaga
     4741 atgttgtggc catgaaagtg aatgacaaga aagacatctc aagattcctg gcatgtaacc
     4801 cagtgatgat cgatgccatc aaggtgtctg ctgctcacag ggcccggtac ttctggggta
     4861 acctacccgg aatgaacagg cccgtgatgg cttcaaagaa tgataagctc gagctgcagg
     4921 actgcctgga gttcagtagg acagcaaagt taaagaaagt gcagacaata accaccaagt
     4981 cgaactccat cagacagggc aaaaaccagc ttttccctgt agtcatgaat ggcaaggacg
     5041 acgttttgtg gtgcactgag ctcgaaagga tcttcggctt ccctgctcac tacacggacg
     5101 tgtccaacat gggccgcggc gcccgtcaga agctgctggg caggtcctgg agtgtaccgg
     5161 tcatcagaca cctgtttgcc cccttgaagg actactttgc ctgtgaatag gcggccgcag
     5221 ttaacgaatt ctctagagga tccagacatg ataagataca ttgatgagtt tggacaaacc
     5281 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5341 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     5401 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     5461 ggtatggctg attatgatcc tgcaagcctc gtcgccgcgg tttattctgt tgacaattaa
     5521 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggga
     5581 tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5641 ttcggctatg actgggcaca acagacgatc ggctgctctg atgccgccgt gttccggctg
     5701 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5761 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5821 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5881 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5941 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6001 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6061 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     6121 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6181 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6241 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6301 ttcctcgtgc tttacggtat cgccgccccc gattcgcagc gcatcgcctt ctatcgcctt
     6361 cttgacgagt tcttctgagc gggactctgg ggttcgaata aagaccgacc aagcgacgtc
     6421 tgagagctcc ctggcgaatt cggtaccaat aaaagagctt tattttcatg atctgtgtgt
     6481 tggtttttgg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6541 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6601 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6661 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6721 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6781 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6841 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6901 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6961 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7021 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7081 aaaaaaggat ctcaagaaga tcctttgatc ttttctagtg tgcg
//
