LOCUS       sequence-320841-        4957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Separately expressing 3 unique sgIGR_dual-MTS (SetI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4957)
  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 4957)
  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-sgIGR_dual-MTS-3x-Set I
FEATURES             Location/Qualifiers
     source          1..4957
                     /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(572..588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    596..612
                     /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        646..894
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1151..1167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1175..1191
                     /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        1225..1473
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1736..1752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1760..1776
                     /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(1828..1846)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1829..1846)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1864..1880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1888..1904
                     /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(1912..1918,1919..1936,1937..1942))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1957..1978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2095..2112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2266..2854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2346..2365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3025..3816,3817..3885))
                     /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     3648..3667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3886..3990)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4058..4076
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4114..4136)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4236..4255
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4322..4777)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4459..4480)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4671..4690
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4903..4925
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4917..4934
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4918..4934
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4941..4957,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4941..4957,1..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 aacaccgcgc tccatgtagt ttgagagcta tgctggaaac agcatagcaa
      361 gttcaaataa ggctagtccg ttatcaactt ggccccggag cagaacgaca ggagttgtgt
      421 ttgtggacga agcaagctca gtcacgacat cacttacgct agcctgcagt ctgctccggg
      481 gccagtggca ccgagtcggt gcttttttta ttggatcgga tgccgggacc gacgagtgca
      541 gaggcgtgca agcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      601 atccgctcac aactcgacaa tgcgtcgaga tccaattagt tattagaggg cctatttccc
      661 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
      721 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
      781 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
      841 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgtggac
      901 accgttggtt tgagagctat gctggaaaca gcatagcaag ttcaaataag gctagtccgt
      961 tatcaacttg gccccggagc agaacgacag gagttgtgtt tgtggacgaa gcaagctcag
     1021 tcacgacatc acttacgcta gcctgcagtc tgctccgggg ccagtggcac cgagtcggtg
     1081 ctttttttat tggatcggat gccgggaccg acgagtgcag aggcgtgcaa gcgagcttgg
     1141 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca actcgacaat
     1201 gcgtcgagat ccaattagtt attagagggc ctatttccca tgattccttc atatttgcat
     1261 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
     1321 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     1381 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     1441 gctttatata tcttgtggaa aggacgaaac accgtctctc gactgttgcg ttgtttgaga
     1501 gctatgctgg aaacagcata gcaagttcaa ataaggctag tccgttatca acttggcccc
     1561 ggagcagaac gacaggagtt gtgtttgtgg acgaagcaag ctcagtcacg acatcactta
     1621 cgctagcctg cagtctgctc cggggccagt ggcaccgagt cggtgctttt tttattggat
     1681 cggatgccgg gaccgacgag tgcagaggcg tgcaagcgag cttggcgtaa tcatggtcat
     1741 agctgtttcc tgtgtgaaat tgttatccgc tcacaactcg agggatccgg gccctctaga
     1801 tgcggccgca tgcataagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc
     1861 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1921 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1981 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2041 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2101 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2161 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2221 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2281 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2341 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2401 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2461 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2521 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2581 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2641 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2701 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2761 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2821 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2881 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2941 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3001 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3061 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3121 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3181 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3241 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3301 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3361 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3421 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3481 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3541 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3601 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3661 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3721 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3781 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3841 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3901 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3961 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4021 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4081 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4141 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4201 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4261 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4321 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4381 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4441 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     4501 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     4561 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     4621 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     4681 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     4741 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     4801 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     4861 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     4921 aaacgacggc cagtgaattg taatacgact cactata
//
