LOCUS       sequence-437324-        4547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid used to place the RNA sequence of interest next to two MS2 
            coat protein recognition sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4547)
  AUTHORS   Bernstein DS, Buter N, Stumpf C, Wickens M
  TITLE     Analyzing mRNA-protein complexes using a yeast three-hybrid system.
  JOURNAL   Methods. 2002 Feb;26(2):123-41. doi: 10.1016/S1046-2023(02)00015-4.
  PUBMED    12054889
REFERENCE   2  (bases 1 to 4547)
  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"; doi: "10"; journalName: 
            "Methods"; date: "2002-02"; volume: "26"; issue: "2"; pages: 
            "123-41"
COMMENT     Sequence Label: pMS2-1
FEATURES             Location/Qualifiers
     source          1..4547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        complement(741..759)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        complement(780..798)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(851..867)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(852..868)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(897..917)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(897..915)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(936..952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(936..952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(949..971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    960..976
                     /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(984..990,991..1008,1009..1014))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1029..1050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1167..1184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1581..2771)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     2400..2419
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(2819..2824,2825..2841,2842..2847))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      complement(2919..3507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        complement(4420..4524)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gccggaattc cggctagaac tagtggatcc cccgggcgat
      721 cctctagagt cgacctgcag acatgggtga tcctcatgtt ttctagagtc gacctgcaga
      781 catgggtgat cctcatgttt tctagagtcg acctgcaggc atgcaagctg gaattcgata
      841 tcaagcttat cgataccgtc gacctcgagg gggggcccgg tacccagctt ttgttccctt
      901 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      961 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     1021 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1081 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1141 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1201 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1261 gataacgcag gaaagaacat gtatgcgtat atataccaat ctaagtctgt gctccttcct
     1321 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca
     1381 aaaaaaaraa waaaaaaaaa atgatgaayy nnsmmmcccc cccccccccc cgatgcgccg
     1441 cgtgcggctg ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc
     1501 attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc
     1561 gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac
     1621 gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg
     1681 ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg
     1741 gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac
     1801 agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg
     1861 aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc
     1921 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     1981 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     2041 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     2101 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     2161 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgacgctc tcccttatgc
     2221 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     2281 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     2341 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     2401 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     2461 atgcgtccgg cgtagaggat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata
     2521 gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct
     2581 ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag
     2641 tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata
     2701 accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg
     2761 ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat
     2821 taaagcttat cgatgataag ctgtcaaaca tgagaattca tgtgagcaaa aggccagcaa
     2881 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2941 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3001 agatactagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3061 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3121 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3181 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3241 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3301 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3361 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3421 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3481 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3541 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3601 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3661 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3721 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3781 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3841 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3901 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3961 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4021 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4081 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4141 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4201 tccgtaagat gcttttctgt gactggtgag tcgttcttcg gggcgaaaac tctcaaggat
     4261 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4321 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4381 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4441 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4501 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
