LOCUS       sequence-409941-        2344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BoxB-petracrRNA in mammalian cells (with a human-U6 
            promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2344)
  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 2344)
  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-BoxB-petracrRNA
FEATURES             Location/Qualifiers
     source          1..2344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        124..372
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     124..144
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     295..314
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    378..396
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     rep_origin      complement(637..1225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(717..736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1396..2187,2188..2256))
                     /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     2019..2038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(2257..2344,1..17))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcgc tagctgtaca
       61 aaaaagcagg ctttaaagga accaattcag tcgactggat ccggtaccaa ggtcgggcag
      121 gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      181 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
      241 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
      301 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      361 ggacgaaaca ccggataggg ccctgaagaa gggccctatc tcttcgcgca agtttaaata
      421 aggctagtcc gttatcaact tgaaaaagtg ggaccgagtc ggtcctctgc catcaaagcg
      481 tgctcagtct gttgacgcgg ttctatctag ttacgcgtta aaccaactag aaattttttt
      541 aagcttgggc cgctcgaggt acctctctac atatgacatg tgagcaaaag gccagcaaaa
      601 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      661 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      721 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      781 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      841 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      901 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      961 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1021 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1081 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1141 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1201 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1261 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1321 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1381 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1441 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1501 cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac cggctccaga
     1561 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1621 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1681 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1741 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1801 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1861 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1921 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1981 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2041 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2101 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2161 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2221 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2281 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2341 taaa
//
