LOCUS       sequence-408317-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses LambdaN-MCP fusion protein in mammalian cells (with a CMV 
            promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  AUTHORS   Choi J, Chen W, Liao H, Li X, Shendure J
  TITLE     A molecular proximity sensor based on an engineered, dual-component 
            guide RNA.
  JOURNAL   Elife. 2025 Feb 12;13:RP98110. doi: 10.7554/eLife.98110.
  PUBMED    39937081
REFERENCE   2  (bases 1 to 5891)
  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: "Elife."; 
            date: "2025-02-12"; volume: "13:RP98110. doi"; pages: " 
            10.7554/eLife.98110"
COMMENT     Sequence Label: pCMV-LambdaN-MCP
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1440..1643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1594..1614
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1640..1664
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1719..1738
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1719..1737
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1745..1754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1754..1783
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1784..1831
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             1856..1921
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1979..1999
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2030..2056
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     polyA_signal    2461..2582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2498..2517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2552..2571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3345..3379
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3870..3886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3870..3886)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3883..3905)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3894..3910
                     /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(3918..3924,3925..3942,3943..3948))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3963..3984
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4101..4118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4272..4860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4352..4371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5031..5822,5823..5891))
                     /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     5654..5673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtattatgcc cagtacatga
     1381 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1441 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1501 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1561 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1621 gggaggttta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     1681 cacgctgttt tgacctccat agaagattct agagctagta atacgactca ctatagggag
     1741 agccgccacc atggagcaga agctgatcag cgaggaagat ctgaagcgcc ccgccgctac
     1801 aaagaaagcc ggccaggcca agaagaagaa aggcggttcc gccagcggcg gcagcatgga
     1861 cgcccaaacc aggagacggg aacggagagc cgagaagcag gctcagtgga aggccgctaa
     1921 ttctggagga tctagcggag gatccaaacg gacagccgac ggaagcgagt tcgagtcacc
     1981 aaagaagaag cggaaagtct ctggcggttc aagcggcggc tctggcggcc ctgctgccaa
     2041 gagggtcaag ttggacggaa gcggagccag caatttcacc cagttcgtgc tggtcgacaa
     2101 cggcggtaca ggcgatgtga ccgtggcccc tagcaacttc gccaacggcg tggccgagtg
     2161 gatcagcagc aacagcagaa gccaggccta taaggtgaca tgcagcgtgc ggcagtcttc
     2221 tgcccagaag cgcaagtaca ccatcaaggt ggaggtgcct aaagtggcta cccaaacagt
     2281 gggaggcgtg gagctgcctg tggcagcctg gcggagctac ctgaacatgg aactcaccat
     2341 ccctatcttc gccacgaaca gcgactgtga gctgatcgtg aaagccatgc agggcctgct
     2401 gaaggacggc aaccccatcc cttctgccat cgccgctaat agtggactgt actaaaacta
     2461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2581 tatcatgtct ggaattgact caaatgatgt caattagtct atcagaagct catctggtct
     2641 cccttccggg ggacaagaca tccctgttta atatttaaac agcagtgttc ccaaactggg
     2701 ttcttatatc ccttgctctg gtcaaccagg ttgcagggtt tcctgtcctc acaggaacga
     2761 agtccctaaa gaaacagtgg cagccaggtt tagccccgga attgactgga ttcctttttt
     2821 agggcccatt ggtatggctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc
     2881 agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc
     2941 cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg
     3001 ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg
     3061 ggctgtccct gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga
     3121 acatgaaata acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa
     3181 tcatgcgtca ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat
     3241 tgacaagcac gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga
     3301 cggattcgcg ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac
     3361 gcagactatc tttctagggt taatctagct gcatcaggat catatcgtcg ggtctttttt
     3421 ccggctcagt catcgcccaa gctggcgcta tctgggcatc ggggaggaag aagcccgtgc
     3481 cttttcccgc gaggttgaag cggcatggaa agagtttgcc gaggatgact gctgctgcat
     3541 tgacgttgag cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg ccatgcacgc
     3601 ctttaacggt gaactgttcg ttcaggccac ctgggatacc agttcgtcgc ggcttttccg
     3661 gacacagttc cggatggtca gcccgaagcg catcagcaac ccgaacaata ccggcgacag
     3721 ccggaactgc cgtgccggtg tgcagattaa tgacagcggt gcggcgctgg gatattacgt
     3781 cagcgaggac gggtatcctg gctggatgcc gcagaaatgg acatggatac cccgtgagtt
     3841 acccggcggg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3901 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3961 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4021 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4081 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4141 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4201 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4261 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4321 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4381 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4441 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4501 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4561 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4621 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4681 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4741 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4801 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4861 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4921 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4981 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5041 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5101 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5161 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5221 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5281 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5341 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5401 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5461 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5521 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5581 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5641 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5701 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5761 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5821 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5881 tgaatactca t
//
