LOCUS       sequence-305959-        3680 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cassette of 10 different binding sites with high affinity to MS2 
            phage coat protein, as extracted from the oligo pool experiment..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3680)
  AUTHORS   Katz N, Tripto E, Granik N, Goldberg S, Atar O, Yakhini Z, Orenstein
            Y, Amit R
  TITLE     Overcoming the design, build, test bottleneck for synthesis of 
            nonrepetitive protein-RNA cassettes.
  JOURNAL   Nat Commun. 2021 Mar 11;12(1):1576. doi: 10.1038/s41467-021-21578-6.
  PUBMED    33707432
REFERENCE   2  (bases 1 to 3680)
  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: "2021-03-11"; volume: "12(1):1576. doi"; pages: " 
            10.1038/s41467-021-21578-6"
COMMENT     Sequence Label: CMV-10xMS2 oligo-library based
FEATURES             Location/Qualifiers
     source          1..3680
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    21..142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(32..51)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     86..105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(395..412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(566..1154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(646..665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1325..2116,2117..2185))
                     /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     1948..1967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2186..2290)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2317..2772
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2404..2423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2614..2635
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2903..2951
                     /note="synthetic polyadenylation signal"
     misc_feature    2965..3056
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3005..3024
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        3123..3426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3427..3630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3581..3601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
       61 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      121 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      181 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
      241 taaggatcga gtatgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
      301 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
      361 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      421 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      481 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      541 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      601 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      661 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      721 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
      781 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      841 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      901 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      961 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1021 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1081 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1141 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1201 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1261 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1321 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1381 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1441 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1501 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1561 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1621 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1681 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1741 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1801 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1861 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1921 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1981 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2041 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2101 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2161 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2281 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     2341 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2401 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2461 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2521 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2581 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2641 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2701 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2761 cgtttacaat ttcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     2821 gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac
     2881 gggaggtact tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     2941 ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa
     3001 caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctggta
     3061 cctagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     3241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     3301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     3361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     3421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     3481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     3541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     3601 gtacggtggg aggtctatat aagcagagct ggtttatgaa ccgtcagatc cgagctccta
     3661 tctagcgtcg cgttgaccat
//
