LOCUS       sequence-292216-        4177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS2V7 cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4177)
  AUTHORS   Tutucci E, Vera M, Biswas J, Garcia J, Parker R, Singer RH
  TITLE     An improved MS2 system for accurate reporting of the mRNA life 
            cycle.
  JOURNAL   Nat Methods. 2017 Nov 13. doi: 10.1038/nmeth.4502.
  PUBMED    29131164
REFERENCE   2  (bases 1 to 4177)
  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: "Nat 
            Methods. 2017 Nov 13. doi: 10.1038/nmeth.4502."
COMMENT     Sequence Label: pET263-pUC57 24xMS2V7
FEATURES             Location/Qualifiers
     source          1..4177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1956..1972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1956..1972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1969..1991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1980..1996
                     /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(2004..2010,2011..2028,2029..2034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2049..2070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2187..2204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2358..2946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2438..2457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3117..3908,3909..3977))
                     /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     3740..3759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3978..4082)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4150..4168
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
      421 tgcatctaga tacccaatgg atcccccctg gcaatcgcgg ggagacgcag gaccaccgcg
      481 tcttcactct cgcttgcgcg gtatacgcag ggagcaacgt cgacacagag gaacaccctg
      541 tgtggcccca cggtgctaca aagaacattc cgtattgctc acatgtggag gaccacccca
      601 caagcgagtc aggaagcatc gcaccattat cccgaacaat cgacacgcag gatcaccgcg
      661 tgggacactc tgttccccgt caaaattgga ccataccgga cgtcatgcag gatcaccgca
      721 tgcatggtgc agaatatcgg catgtctact cgtacagtca gtgtggtcag gaccaccgac
      781 cacttctatt ctattcatct tttcggttgt gcaggtattc acgagaagac gatcacgctt
      841 ctcgactacc ttcatcatac atggtgtgca gatgtttttt gccaatggag gaacacccca
      901 ttctcaagtt tacactctgc tatggcagca ctgtcgcaga ggaacaccct gcgagccccc
      961 ggtgcgtgta ttgcatctgc cttgcgagca ttcacaggga cgaacacgcc ctgccgttgc
     1021 attacttcaa tatgggtgct ctgtcgtcgt catttgcgca ggaccaccgc gcatatatca
     1081 tcagcactcg tgcggatact tctgggattc cgcgagctca ggaacaccga gctctggcga
     1141 cagagaccct cacacagatc ccccctggca atcgcgggga gacgcaggac caccgcgtct
     1201 tcactctcgc ttgcgcggta tacgcaggga gcaacgtcga cacagaggaa caccctgtgt
     1261 ggccccacgg tgctacaaag aacattccgt attgctcaca tgtggaggac caccccacaa
     1321 gcgagtcagg aagcatcgca ccattatccc gaacaatcga cacgcaggat caccgcgtgg
     1381 gacactctgt tccccgtcaa aattggacca taccggacgt catgcaggat caccgcatgc
     1441 atggtgcaga atatcggcat gtctactcgt acagtcagtg tggtcaggac caccgaccac
     1501 ttctattcta ttcatctttt cggttgtgca ggtattcacg agaagacgat cacgcttctc
     1561 gactaccttc atcatacatg gtgtgcagat gttttttgcc aatggaggaa caccccattc
     1621 tcaagtttac actctgctat ggcagcactg tcgcagagga acaccctgcg agcccccggt
     1681 gcgtgtattg catctgcctt gcgagcattc acagggacga acacgccctg ccgttgcatt
     1741 acttcaatat gggtgctctg tcgtcgtcat ttgcgcagga ccaccgcgca tatatcatca
     1801 gcactcgtgc ggatacttct gggattccgc gagctcagga acaccgagct ctggcgacag
     1861 agaccctcac acagatctat cgatctcgag attgggtcat cggatcccgg gcccgtcgac
     1921 tgcagaggcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1981 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2041 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2101 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2161 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2221 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2281 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2341 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2401 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2461 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2521 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2581 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2701 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2761 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2821 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2881 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2941 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3001 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4081 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     4141 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
