LOCUS       sequence-429613-        7270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MS2 coat protein (MCP) fused to YPet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7270)
  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 7270)
  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: MCP-YPet
FEATURES             Location/Qualifiers
     source          1..7270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             52..285
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(404..422)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        490..594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(595..663,664..1455)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(813..832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1626..2214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2115..2134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2368..2385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2625..2646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2661..2666,2667..2684,2685..2691)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2699..2715
                     /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)."
     primer_bind     2704..2726
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2723..2739
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2723..2739
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2789..2808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3000..3633
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3680..3805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4302..4535
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4720..4764
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5057..5174
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5264..5475
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5521..5530
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5905..5907,5908..5910,5911..6621)
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDSNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     primer_bind     complement(6211..6230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             6628..6648
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    6659..7247
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6712..6732)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7130..7141)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
       61 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
      121 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
      181 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
      241 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
      301 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcccg
      361 ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt gatacgccta
      421 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
      481 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      541 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      601 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      661 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      721 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      781 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
      841 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      901 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
      961 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1021 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1081 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1141 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1201 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1261 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1321 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1381 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1441 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1501 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1741 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2281 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2341 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2401 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag caagctcatg
     2461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2521 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgtggcac
     2581 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     2641 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     2701 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttcacaaat
     2761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2821 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     2881 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     2941 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt
     3001 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
     3061 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     3121 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     3181 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     3241 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     3301 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     3361 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     3421 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     3481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     3541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     3601 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     3661 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     3721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     3781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     3841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     3901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     3961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     4021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     4081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     4141 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     4201 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     4261 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     4321 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     4381 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     4441 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     4501 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     4561 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     4621 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     4681 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     4741 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     4801 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     4861 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     4921 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     4981 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccgaat tgacaaatgg
     5041 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     5101 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     5161 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg actagcgatg
     5221 ctagctgatg cgggcccttc gagtggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     5281 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     5341 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     5401 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     5461 gccgccagaa cacaggtgtc gtgacgcggt tccggttaag gtgaacgcgt gcaggctggc
     5521 gccaccatgg cttctaactt tactcagttc gttctcgtcg acaatggcgg aactggcgac
     5581 gtgactgtcg ccccaagcaa cttcgctaac gggatcgctg aatggatcag ctctaactcg
     5641 cgttcacagg cttacaaagt aacctgtagc gttcgtcaga gctctgcgca gaatcgcaaa
     5701 tacaccatca aagtcgaggt gcctaaaggc gcctggcgtt cgtacttaaa tatggaacta
     5761 accattccaa ttttcgccac gaattccgac tgcgagctta ttgttaaggc aatgcaaggt
     5821 ctcctaaaag atggaaaccc gattccctca gcaatcgcag caaactccgg catctacggg
     5881 ggatctggta gcggcgctgg atccatggtg agcaaaggcg aagagctgtt caccggcgtg
     5941 gtgcccatcc tggtggagct ggacggcgac gtgaacggcc acaagttcag cgtgagcggc
     6001 gagggcgagg gcgacgccac ctacggcaag ctgaccctga agctgctgtg caccaccggc
     6061 aagctgcccg tgccctggcc caccctggtg accaccctgg gctacggcgt gcagtgcttc
     6121 gcccggtacc ccgaccacat gaagcagcac gacttcttca agagcgccat gcccgagggc
     6181 tacgtgcagg agcggaccat cttcttcaag gacgacggca actacaagac ccgggccgag
     6241 gtgaagttcg agggcgacac cctggtgaac cggatcgagc tgaagggcat cgacttcaag
     6301 gaggacagca acatcctggg ccacaagctg gagtacaact acaacagcca caacgtgtac
     6361 atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg gcacaacatc
     6421 gaggacggcg gcgtgcagct ggccgaccac taccagcaga acacccccat cggcgacggc
     6481 cccgtgctgc tgcccgataa ccactacctg agctaccaga gcgccctgtt caaggacccc
     6541 aacgagaagc gggaccacat ggtgctgctg gagttcctga ccgccgccgg catcacagag
     6601 ggcatgaacg agctgtataa gctcgagcca aagaaaaagc ggaaagtgta atctagataa
     6661 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     6721 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     6781 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     6841 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     6901 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     6961 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     7021 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     7081 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     7141 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     7201 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta
     7261 agaccaatga
//
