LOCUS       sequence-409944-        2284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses crRNA-MS2 in mammalian cells (with a human-U6 promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2284)
  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 2284)
  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: pU6-crRNA-MS2
FEATURES             Location/Qualifiers
     source          1..2284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      104..692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     593..612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_RNA        complement(840..858)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        complement(897..1145)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(955..974)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1125..1145)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        1252..1356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1357..1425,1426..2217)
                     /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     complement(1575..1594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
       61 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
      121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      721 tcctggcctt ttgctggcct tttgctcaca tgtcatatgt agagaggtac ctcgagcggc
      781 ccaagcttaa aaaaatttct agttggttta acgcgtaact agatagaacc gcgtcaacca
      841 catgggtgat cctcatgtgg agaagcgctc ttaaactcac gtgctcagtc tgggccggtg
      901 tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca agttacggta
      961 agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat
     1021 tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcta
     1081 attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg
     1141 ccctcttcct gcccgacctt ggtaccggat ccagtcgact gaattggttc ctttaaagcc
     1201 tgcttttttg tacagctagc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1261 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1321 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1381 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1441 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1501 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1561 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1621 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1681 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1741 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1801 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1861 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1921 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1981 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2041 attgctgata aatctggagc cggtgagcgt ggatctcgcg gtatcattgc agcactgggg
     2101 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2161 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2221 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2281 agga
//
