LOCUS       sequence-311813-        4116 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4116)
  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 4116)
  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..4116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    103..151
                     /note="synthetic polyadenylation signal"
     misc_feature    165..256
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     205..224
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     enhancer        323..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1337..1458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1348..1367)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1402..1421
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1711..1728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1882..2470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1962..1981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2641..3432,3433..3501))
                     /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     3264..3283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3502..3606)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3633..4088
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3720..3739)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3930..3951
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata
       61 agtaagtaat attaaggtac gggaggtact tggagcggcc gcaataaaat atctttattt
      121 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat
      181 caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt
      241 gccagaacat ttctctggta cctagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttatgaa
      841 ccgtcagatc cgagctccta tctagcgtcg cgttgacgaa ttcttacatg aagacgatta
      901 cgcttcacgc ctgctggtcg tgacagttga ccattaggca actcgagaca atgctacctt
      961 accgtcggcc cacttgtttt tacccatgag aacacgatta cggttctcgc ctgctggtcg
     1021 tgacacatga ggattaccca tgtgaaacag ccgtcgcctt gaagccgaga acaacttgac
     1081 cattaggcaa gtatggattg ggtctccaat tcctgtctgt agatcaccat tagggatctc
     1141 atcaggcttc gcagtatgca acgcttgcga tgcgaggatt acccgcattt caccgctttc
     1201 ctaagagttc agcattagcg aactgtacta actcgcagtt gtgaggagta cccacaacag
     1261 aaacgtcacg tcctggcaca tgaggattac ccatgtggaa gaattccccc taagcttccg
     1321 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     1381 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     1441 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     1501 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     1561 gatcgagtat gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     1621 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     1681 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3481 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3541 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3601 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     3661 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3721 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3781 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3841 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3901 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3961 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4021 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4081 tacaatttgc cattcgccat tcaggctgcg caactg
//
