LOCUS       sequence-431994-        4545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid allows you 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 4545)
  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 4545)
  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: pMS2-2
FEATURES             Location/Qualifiers
     source          1..4545
                     /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        749..767
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        788..806
                     /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(1579..2769)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     2398..2417
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(2817..2822,2823..2839,2840..2845))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      complement(2917..3505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2997..3016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        complement(4418..4522)
                     /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 cccgggcagc
      721 ttgcatgcct gcaggtcgac tctagaaaac atgaggatca cccatgtctg caggtcgact
      781 ctagaaaaca tgaggatcac ccatgtctgc aggtcgactc tagaggatcg gaattcgata
      841 tcaagcttat cgataccgtc gacctcgagg gggggcccgg tacccagctt ttgttccctt
      901 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      961 tgttatccgc tcacaattcc acacaacata cgagccggaa 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 tttcaaagaa accgaaatca
     1381 aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaag gggggggggg atgcgccgcg
     1441 tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat
     1501 tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga
     1561 ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc
     1621 ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct
     1681 cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca gtgatcgaag ttaggctggt
     1741 aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag
     1801 catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa
     1861 ggccatccag cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat
     1921 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     1981 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     2041 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     2101 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     2161 gaaggagctg actgggttga aggctctcaa gggcatcggt cgacgctctc ccttatgcga
     2221 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag
     2281 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     2341 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     2401 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     2461 gcgtccggcg tagaggatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt
     2521 ggctccaagt agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc
     2581 gagaacgggt gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg
     2641 actggcgatg ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac
     2701 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     2761 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     2821 aagcttatcg atgataagct gtcaaacatg agaattcatg tgagcaaaag gccagcaaaa
     2881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3541 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3601 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3661 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3721 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3781 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3841 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3901 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3961 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4021 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4081 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4141 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4201 cgtaagatgc ttttctgtga ctggtgagtc gttcttcggg gcgaaaactc tcaaggatct
     4261 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4321 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4381 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4441 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4501 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
