LOCUS       sequence-315349-        4978 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 4978)
  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 4978)
  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..4978
                     /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(1169..1185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1193..1209
                     /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        1243..1491
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1757..1773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1781..1797
                     /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(1849..1867)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1850..1867)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1885..1901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1909..1925
                     /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(1933..1939,1940..1957,1958..1963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1978..1999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2116..2133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2287..2875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2367..2386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3046..3837,3838..3906))
                     /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     3669..3688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3907..4011)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4079..4097
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4135..4157)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4257..4276
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4343..4798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4480..4501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4692..4711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4924..4946
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4938..4955
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4939..4955
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4962..4978,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4962..4978,1..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 gcgtgcaagc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1201 cgctcacaac tcgacaatgc gtcgagatcc aattagttat tagagggcct atttcccatg
     1261 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
     1321 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     1381 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     1441 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cgtacaccct
     1501 tgtgtacaga gctgtttgag agctatgctg gaaacagcat agcaagttca aataaggcta
     1561 gtccgttatc aacttggccc cggagcagaa cgacaggagt tgtgtttgtg gacgaagcaa
     1621 gctcagtcac gacatcactt acgctagcct gcagtctgct ccggggccag tggcaccgag
     1681 tcggtgcttt ttttattgga tcggatgccg ggaccgacga gtgcagaggc gtgcaagcga
     1741 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaactc
     1801 gagggatccg ggccctctag atgcggccgc atgcataagc ttgagtattc tatagtgtca
     1861 cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     1921 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1981 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2041 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2101 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2161 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2221 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2281 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2341 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2401 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2461 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2521 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2581 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2641 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2701 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2761 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2821 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2881 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2941 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3001 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3061 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3121 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3181 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3241 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3301 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3361 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3421 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3481 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3541 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3601 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3661 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3721 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3781 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3841 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3901 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3961 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4021 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4081 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4141 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4201 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4261 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4321 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     4381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     4561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     4621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     4681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     4801 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4861 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     4921 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactata
//
