LOCUS       sequence-429728-        4821 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses binding-deficient W42A mutant of 2xHP1beta chromo domain 
            fused to C-terminal part of IFP2.0 as negative control for H3K9me2/3
            detection in BiAD sensors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4821)
  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 4821)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: 2xHP1bCD_W42A-IFP2.0C
FEATURES             Location/Qualifiers
     source          1..4821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     237..256
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(345..702)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      359..494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        511..702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     659..679
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(704..808)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      835..1290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(922..941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1132..1153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1297..1418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1308..1327)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1362..1381
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    1542..1607
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2411..2420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2459..2662)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2488..2508)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2663..2966
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3141..3729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3221..3240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4058..4105
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4130..4149
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(4337..4821,1..310))
                     /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             complement(join(4337..4821,1..310))
                     /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(4448..4467)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
       61 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
      121 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
      181 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
      241 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
      301 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
      361 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
      421 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
      481 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
      541 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
      601 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      661 cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc ctttttcaat
      721 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
      781 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
      841 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      901 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      961 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1021 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1141 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1201 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1261 attttaacaa aatattaacg cttacaattt acgcgttaag atacattgat gagtttggac
     1321 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1381 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1441 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1501 aatgtggtat ggctgattat gatcagttat ctagatccgg tggatcccgg gcccgcggta
     1561 ccgtcgactg cagaattcga agcttgagct cgagatctga gtccggatta ggcttctttc
     1621 ctctgcacct gcagggacag gagcctcccc agatactcca gggtggtccg cagatcgggt
     1681 ggcagcacgt agggtgtctg atggtggcag acaatcagtc cccacagctg gccgccgacc
     1741 acgacgctga cgctcaggga ggagcccact cccatattcc tcaggtactg catgtgcatt
     1801 ggggatgtag ctctgaggac agctccgccc agaggtgtag gagcatttgt ctggggattc
     1861 agcacggggt cgaggggcac tgctgcagcc ctggtgtctg cggtcagccg gagcaggtgt
     1921 cttgtgtaga gggccctagc ttgtgcaggg gtggtggatg cgggaaacct atgcccgagg
     1981 taagcctgca tgccctcgcg ccttgcttcg gcaataacct ctcctgttgc gtctggtgca
     2041 aacttgtaca gcatcacccg gtcgaaacct gacagttccc ggactgtttg ggttgcgact
     2101 tcagccagtg cccggagatt aggagcactc tccagagcga acattgcatt tctcagagcg
     2161 tggggtccaa tagagtccgt gccctcctgg ccttccgacg tggatgaacc cgaggtttgt
     2221 gacgatccag tctcatgtgc tgttttctgt gactgtagaa actcagcaat aaggtcaggg
     2281 caatccagat tctcttctgg ctcccaagtg ttgtcctcat ctgagaaacc cttcgccttt
     2341 agaagatatt ccaccttgcc cttgacaact cgccgatcaa gaactttttc caccacatat
     2401 tcctcttcct ccatggtggc gaccggtagc gctagcggat ctgacggttc actaaaccag
     2461 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     2521 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     2581 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     2641 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     2701 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     2761 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     2821 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     2881 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     2941 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     3001 ccgtaattga ttactattaa taactaatgc atggcggtaa tacggttatc cacagaatca
     3061 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3121 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3181 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3241 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3301 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3361 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3421 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3481 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3541 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3601 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3661 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3721 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3781 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3841 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca
     3901 gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg
     3961 ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg
     4021 acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc
     4081 gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc
     4141 cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc
     4201 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     4261 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     4321 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     4381 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     4441 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     4501 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4561 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     4621 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     4681 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     4741 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     4801 cggcaggagc aaggtgagat g
//
