LOCUS       sequence-320822-        3799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing sgControl_dual-MTS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3799)
  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 3799)
  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-sgControl_dual-MTS
FEATURES             Location/Qualifiers
     source          1..3799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..21
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2..20
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        86..334
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     86..106
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     257..276
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(596..612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    620..636
                     /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(688..706)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(689..706)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(724..740)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    748..764
                     /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(772..778,779..796,797..802))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    817..838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(955..972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1126..1714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1206..1225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1885..2676,2677..2745))
                     /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     2508..2527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2746..2850)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2918..2936
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2974..2996)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3096..3115
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3182..3637)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3319..3340)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3531..3550
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3763..3785
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3777..3794
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3778..3794
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtaatacgac tcactatagg gcgaattggc caagtcggcc gagctcgaat tcgtcgacaa
       61 tgcgtcgaga tccaattagt tattagaggg cctatttccc atgattcctt catatttgca
      121 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
      181 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
      241 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
      301 ggctttatat atcttgtgga aaggacgaaa caccgagctg cacgctgccg tcgtttgaga
      361 gctatgctgg aaacagcata gcaagttcaa ataaggctag tccgttatca acttggcccc
      421 ggagcagaac gacaggagtt gtgtttgtgg acgaagcaag ctcagtcacg acatcactta
      481 cgctagcctg cagtctgctc cggggccagt ggcaccgagt cggtgctttt tttattggat
      541 cggatgccgg gaccgacgag tgcagaggcg tgcaagcgag cttggcgtaa tcatggtcat
      601 agctgtttcc tgtgtgaaat tgttatccgc tcacaactcg agggatccgg gccctctaga
      661 tgcggccgca tgcataagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc
      721 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      781 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      841 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      901 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      961 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1021 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1081 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     1501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     2401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     2461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     2521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2761 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2821 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     2881 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     2941 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3001 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     3061 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     3121 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3181 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3241 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3301 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3361 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3421 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3481 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3541 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3601 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     3661 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     3721 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     3781 aaacgacggc cagtgaatt
//
