LOCUS       sequence-429612-        6456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MBD1 MBD domain fused to C-terminal part of IFP2.0 for 5mC
            detection in BiAD sensors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6456)
  AUTHORS   Kohler AR, Hausser J, Harsch A, Bernhardt S, Haussermann L, Brenner 
            LM, Lungu C, Olayioye MA, Bashtrykov P, Jeltsch A
  TITLE     Modular dual-color BiAD sensors for locus-specific readout of 
            epigenome modifications in single cells.
  JOURNAL   Cell Rep Methods. 2024 Mar 26:100739. doi: 
            10.1016/j.crmeth.2024.100739.
  PUBMED    38554702
REFERENCE   2  (bases 1 to 6456)
  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: "Cell Rep 
            Methods. 2024 Mar 26:100739. doi: 10.1016/j.crmeth.2024.100739."
COMMENT     Sequence Label: MBD-IFP2.0C
FEATURES             Location/Qualifiers
     source          1..6456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     875..894
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        875..893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             989..1054
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(2021..2039)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2022..2039)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2059..2076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2065..2289
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2335..2763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2422..2441)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2632..2653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2772..2792)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2777..3106
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2957..3092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3019..3038
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3173..3967
                     /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(3227..3246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3837..3856
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4141..4262
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4178..4197)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4232..4251
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4311..4327)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4311..4327)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4324..4346)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4335..4351
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4359..4365,4366..4383,4384..4389))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4404..4425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4542..4559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4713..5301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4793..4812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5472..6263,6264..6332))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6095..6114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6333..6437)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg ataaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      841 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      901 aagcttctcg ggcccaataa tcattttatt ttgactgata gtgacctgtt cgttgcaaca
      961 aattgatgag caatgctttt ttataatgga ctataaggac cacgacggag actacaagga
     1021 tcatgatatt gattacaaag acgatgacga taagatggct gaggactggc tggactgccc
     1081 ggccctgggc cctggctgga agcgccgcga agtctttcgc aagtcagggg ccacctgtgg
     1141 acgctcagac acctattacc agagccccac aggagacagg atccgaagca aagttgagct
     1201 gactcgatac ctgggccctg cgtgtgatct caccctcttc gacttcaaac aaggcatctt
     1261 gtgctatcca gcccccaagg cccatcccgt ggcggttgcc agcaagaagc gaaagaagcc
     1321 ttcaaggcca gccaagactc ggaaacgtca ggttggaccc cagagtggtg aggtcaggaa
     1381 ggaggccccg atcgatggtg gcggtggctc tggaggtggt gggtcctccg gagactctat
     1441 tggaccccac gctctgagaa atgcaatgtt cgctctggag agtgctccta atctccgggc
     1501 actggctgaa gtcgcaaccc aaacagtccg ggaactgtca ggtttcgacc gggtgatgct
     1561 gtacaagttt gcaccagacg caacaggaga ggttattgcc gaagcaaggc gcgagggcat
     1621 gcaggcttac ctcgggcata ggtttcccgc atccaccacc cctgcacaag ctagggccct
     1681 ctacacaaga cacctgctcc ggctgaccgc agacaccagg gctgcagcag tgcccctcga
     1741 ccccgtgctg aatccccaga caaatgctcc tacacctctg ggcggagctg tcctcagagc
     1801 tacatcccca atgcacatgc agtacctgag gaatatggga gtgggctcct ccctgagcgt
     1861 cagcgtcgtg gtcggcggcc agctgtgggg actgattgtc tgccaccatc agacacccta
     1921 cgtgctgcca cccgatctgc ggaccaccct ggagtatctg gggaggctcc tgtccctgca
     1981 ggtgcagagg aaagaagcct aataatctag agggccctat tctatagtgt cacctaaatg
     2041 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2101 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2161 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2221 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2281 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2341 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2401 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2461 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2521 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2581 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2641 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2701 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2761 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2881 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2941 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3001 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3061 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3121 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3181 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3241 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3301 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3361 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3421 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3481 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3541 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3601 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3661 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3721 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3781 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3841 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3901 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3961 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4021 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4081 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4141 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4201 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4261 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4321 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4381 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4441 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4501 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4561 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4621 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4681 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4741 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4801 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4861 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4921 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4981 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5041 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5101 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5161 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5221 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5281 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5341 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5401 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5461 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5521 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5581 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5641 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5701 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5761 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5821 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5881 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5941 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6001 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6061 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6121 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6181 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6241 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6301 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6361 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6421 caaatagggg ttccgcgcac atttccccga aaagtg
//
