LOCUS       sequence-269561-        5150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of MS2BS-RNAsite hybrid. Homology regions for 
            recombination with pAWH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5150)
  AUTHORS   Yang JS, Garriga-Canut M, Link N, Carolis C, Broadbent K, 
            Beltran-Sastre V, Serrano L, Maurer SP
  TITLE     rec-YnH enables simultaneous many-by-many detection of direct 
            protein-protein and protein-RNA interactions.
  JOURNAL   Nat Commun. 2018 Sep 14;9(1):3747. doi: 10.1038/s41467-018-06128-x.
  PUBMED    30217970
REFERENCE   2  (bases 1 to 5150)
  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 
            Commun."; date: "2018-09-14"; volume: "9(1):3747. doi"; pages: " 
            10.1038/s41467-018-06128-x"
COMMENT     Sequence Label: pMS22H
FEATURES             Location/Qualifiers
     source          1..5150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      7..54
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(136..160)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      complement(597..1185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(677..696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1514..1561
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1586..1605
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1793..2587)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1793..2587)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1904..1923)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2514..2533
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2588..2692)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2735..2753
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2890..3171
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3172..3846
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(3949..4404)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4086..4107)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4298..4317
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4431..4837
                     /gene="RPR1"
                     /label=RPR1 promoter
                     /note="promoter for the S. cerevisiae RNase P RNA gene"
     misc_RNA        4902..4920
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        4941..4959
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      4992..5054
                     /label=RPR1 terminator
                     /note="transcription terminator for the S. cerevisiae RNase
                     P RNA gene"
ORIGIN
        1 gcataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgcgcgct
       61 tgcagccaag ctaattccgg gcgaatttct tatgatttat gatttttatt attaaataag
      121 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgttatc
      181 cctagtttaa acacccataa tacccataat agctgtttgc caattaccct gttatcccta
      241 ttacttctaa atctgaggag gaaaatcgaa tcgctacctt agcgcctgtt tcaggctgtc
      301 tatgtgtgac tgttgagctg taacaagttg tctcaggtgt tcaatttcat gttctagttg
      361 ctttgtttta ctggtttcac ctgttctatt aggtgttaca tgctgttcat ctgttacatt
      421 gtcgatctgt tcatggtgaa caggcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      481 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      541 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      601 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      661 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      721 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      781 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      841 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      901 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      961 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1021 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1081 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1141 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1201 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1261 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1321 atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc gacgttggct
     1381 gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga agaaacgcgg
     1441 gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc ctgggaccga
     1501 accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct ttttattgcc
     1561 gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccta
     1621 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     1681 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     1741 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
     1801 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     1861 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     1921 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     1981 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgagtc acgacgagat
     2041 cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     2101 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     2161 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     2221 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     2281 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     2341 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     2401 cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     2461 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     2521 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     2581 caatcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2641 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatcta
     2701 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2761 tcttcactta tccctattac ttctaaatct gaggaggaaa atcgaatcgc taccttagga
     2821 ccgttatagt tactatcacc atatatgtaa ctggtgattc tttcgctacc ttagagtgca
     2881 ccatagatca acgacattac tatatatata atataggaag catttaatag aacagcatcg
     2941 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
     3001 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
     3061 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
     3121 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
     3181 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
     3241 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
     3301 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
     3361 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
     3421 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
     3481 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
     3541 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
     3601 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
     3661 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
     3721 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
     3781 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
     3841 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg
     3901 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     3961 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4021 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4081 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4141 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4201 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4261 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4321 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4381 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcggatcc gatctgccaa
     4441 ttgaacataa catggtagtt acatatacta gtaatatggt tcggcacaca ttaaaagtat
     4501 aaaaactatc tgaattacga attacatata ttggtcataa aaatcaatca atcatcgtgt
     4561 gttttatatg tctcttatct aagtataaga atatccatag ttaatattca cttacgctac
     4621 cttttaacct gtaatcattg tcaacaggat atgttaacga cccacattga taaacgctag
     4681 tatttctttt tcctcttctt attggccggc tgtctctata ctcccctata gtctgtttct
     4741 tttcgtttcg attgttttac gtttgaggcc tcgtggcgca catggtacgc tgtggtgctc
     4801 gcggctggga acgaaactct gggagctgcg attggcagaa ttccggctag aactagtgga
     4861 tcccccgggc agcttgcatg cctgcaggtc gactctagaa aacatgagga tcacccatgt
     4921 ctgcaggtcg actctagaaa acatgaggat cacccatgtc tgcaggtcga ctctagagga
     4981 tcggaattcc cccatatcca acttccaatt taatctttct tttttaattt tcacttattt
     5041 gcgatacaga aagaaaaaag cgatagtaac tattgaattt tgtttggatt tggttagatt
     5101 agatatggtt tctctttata tttacatgct aaaaatgggc tacaccagag
//
