LOCUS       sequence-320842-        4979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Separately expressing 3 unique sgMUC4_dual-MTS (SetI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4979)
  AUTHORS   Mao S, Ying Y, Wu X, Krueger CJ, Chen AK
  TITLE     CRISPR/dual-FRET molecular beacon for sensitive live-cell imaging of
            non-repetitive genomic loci.
  JOURNAL   Nucleic Acids Res. 2019 Nov 18;47(20):e131. doi: 10.1093/nar/gkz752.
  PUBMED    31504824
REFERENCE   2  (bases 1 to 4979)
  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: "Nucleic 
            Acids Res."; date: "2019-11-18"; volume: "47(20):e131. doi"; pages: 
            " 10.1093/nar/gkz752"
COMMENT     Sequence Label: pGEM-sgMUC4_dual-MTS-3x-Set I
FEATURES             Location/Qualifiers
     source          1..4979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        68..316
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(582..598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    606..622
                     /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        656..904
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1170..1186)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1194..1210
                     /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        1244..1492
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1758..1774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1782..1798
                     /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(1850..1868)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1851..1868)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1886..1902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1910..1926
                     /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(1934..1940,1941..1958,1959..1964))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1979..2000
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2117..2134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2288..2876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2368..2387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3047..3838,3839..3907))
                     /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     3670..3689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3908..4012)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4080..4098
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4136..4158)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4258..4277
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4344..4799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4481..4502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4693..4712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4925..4947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4939..4956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4940..4956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4963..4979,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4963..4979,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg gccaagtcgg ccgagctcga attcgtcgac aatgcgtcga gatccaatta
       61 gttattagag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
      121 tagagagata attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
      181 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
      241 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
      301 gaaaggacga aacaccgaag agtggaggcc gtgcgcgggt ttgagagcta tgctggaaac
      361 agcatagcaa gttcaaataa ggctagtccg ttatcaactt ggccccggag cagaacgaca
      421 ggagttgtgt ttgtggacga agcaagctca gtcacgacat cacttacgct agcctgcagt
      481 ctgctccggg gccagtggca ccgagtcggt gcttttttta ttggatcgga tgccgggacc
      541 gacgagtgca gaggcgtgca agcgagcttg gcgtaatcat ggtcatagct gtttcctgtg
      601 tgaaattgtt atccgctcac aactcgacaa tgcgtcgaga tccaattagt tattagaggg
      661 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
      721 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
      781 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
      841 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
      901 caccgcaagc aagggaagcg acaagggttt gagagctatg ctggaaacag catagcaagt
      961 tcaaataagg ctagtccgtt atcaacttgg ccccggagca gaacgacagg agttgtgttt
     1021 gtggacgaag caagctcagt cacgacatca cttacgctag cctgcagtct gctccggggc
     1081 cagtggcacc gagtcggtgc tttttttatt ggatcggatg ccgggaccga cgagtgcaga
     1141 ggcgtgcaag cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1201 ccgctcacaa ctcgacaatg cgtcgagatc caattagtta ttagagggcc tatttcccat
     1261 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     1321 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     1381 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     1441 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgtacaccc
     1501 ttgtgtacag agctgtttga gagctatgct ggaaacagca tagcaagttc aaataaggct
     1561 agtccgttat caacttggcc ccggagcaga acgacaggag ttgtgtttgt ggacgaagca
     1621 agctcagtca cgacatcact tacgctagcc tgcagtctgc tccggggcca gtggcaccga
     1681 gtcggtgctt tttttattgg atcggatgcc gggaccgacg agtgcagagg cgtgcaagcg
     1741 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaact
     1801 cgagggatcc gggccctcta gatgcggccg catgcataag cttgagtatt ctatagtgtc
     1861 acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1921 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     1981 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2041 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2101 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2161 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2221 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2281 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2341 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2401 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2461 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2521 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2581 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2641 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2701 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2761 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2821 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2881 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2941 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3001 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3061 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3181 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3901 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4021 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4081 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     4141 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     4201 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     4261 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     4321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     4381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     4861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     4921 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactata
//
