LOCUS       sequence-310337-        4218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4xMS2 stem-loop-hTR-500 bp genomic sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4218)
  AUTHORS   Han S, Zhao BS, Myers SA, Carr SA, He C, Ting AY
  TITLE     RNA-protein interaction mapping via MS2- or Cas13-based APEX 
            targeting.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Aug 24. pii: 2006617117. doi: 
            10.1073/pnas.2006617117.
  PUBMED    32839320
REFERENCE   2  (bases 1 to 4218)
  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: "Proc Natl 
            Acad Sci U S A. 2020 Aug 24. pii: 2006617117. doi: 
            10.1073/pnas.2006617117."
COMMENT     Sequence Label: 4xMS2-hTR
FEATURES             Location/Qualifiers
     source          1..4218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(1740..1756)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1740..1756)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1753..1775)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1764..1780
                     /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(1788..1794,1795..1812,1813..1818))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1833..1854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1971..1988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2142..2730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2222..2241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2901..3692,3693..3761))
                     /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     3524..3543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3762..3866)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3956..3975
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4169..4191
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4183..4200
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4184..4200
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      601 acccactgct tactggcgca cgagcatcag ccgtgctctc aggtctaatc ttcaatcgac
      661 gacgatcacg cgtcgctatt aatcccaaca aacttcgtac accatcaggg tacgtctcag
      721 gtctaatctt caatcagaca ccatcagggt ctgtcacttg ggttgcggag ggtgggcctg
      781 ggaggggtgg tggccatttt ttgtctaacc ctaactgaga agggcgtagg cgccgtgctt
      841 ttgctccccg cgcgctgttt ttctcgctga ctttcagcgg gcggaaaagc ctcggcctgc
      901 cgccttccac cgttcattct agagcaaaca aaaaatgtca gctgctggcc cgttcgcccc
      961 tcccggggac ctgcggcggg tcgcctgccc agcccccgaa ccccgcctgg aggccgcggt
     1021 cggcccgggg cttctccgga ggcacccact gccaccgcga agagttgggc tctgtcagcc
     1081 gcgggtctct cgggggcgag ggcgaggttc aggcctttca ggccgcagga agaggaacgg
     1141 agcgagtccc cgcgcgcggc gcgattccct gagctgtggg acgtgcaccc aggactcggc
     1201 tcacacatgc agttcgcttt cctgttggtg gggggaacgc cgatcgtgcg catccgtcac
     1261 ccctcgccgg caatgggggc ttgtgaaccc ccaaacctga ctgactgggc cagtgtgctg
     1321 caaattggca ggagacgtga aggcacctcc aaagtcggcc aaaatgaatg ggcagtgagc
     1381 cggggttgcc tggagccgtt cctgcgtggg ttctcccgtc ttccgctttt tgttgccttt
     1441 tatggttgta ttacaactta gttcctgctc tgcagatttt gttgaggttt ttgcttctcc
     1501 caaggtagat ctcgaccagt cccctcaacg gggtgtggga gaacagtcat ttttttttga
     1561 gagatcattt aacatttaat gaatatttaa ttagaagatc taaatgaaca ttggaaattg
     1621 tgttccttta atggtcatcg gtttatgcca gaggttagaa gtttcttttt tgaaaaatta
     1681 gaccttggcg atgaccttga gcagtaggat ataaccccca caagcttggc gtaatcatgg
     1741 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1801 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1861 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1921 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1981 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2041 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2101 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2161 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2221 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2281 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2341 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2401 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2461 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2521 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2581 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2641 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2701 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2761 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2821 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2881 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2941 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3001 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3061 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3121 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3181 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3241 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3301 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3361 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3421 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3481 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3541 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3601 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3661 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3721 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3781 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3841 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3901 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag ctatgcggca
     3961 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     4021 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     4081 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     4141 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     4201 gaattcgagc tcggtact
//
