LOCUS       sequence-324347-        2358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB1 encoding a MS2 binding protein with an additional SV40 NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2358)
  AUTHORS   Baumschabl M, Prielhofer R, Mattanovich D, Steiger MG
  TITLE     Fine-Tuning of Transcription in Pichia pastoris Using dCas9 and RNA 
            Scaffolds.
  JOURNAL   ACS Synth Biol. 2020 Dec 18;9(12):3202-3209. doi: 
            10.1021/acssynbio.0c00214. Epub 2020 Nov 12.
  PUBMED    33180466
REFERENCE   2  (bases 1 to 2358)
  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: 
            "10.1021/acssynbio.0c00214"; journalName: "ACS Synth Biol"; date: 
            "2020-12-18- 18"; volume: "9"; issue: "12"; pages: "3202-3209"
COMMENT     Sequence Label: BB1_23_MS2bind_only_SV40NLS
FEATURES             Location/Qualifiers
     source          1..2358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     610..629
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(703..805)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(806..837)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     885..901
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     885..901
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             988..1008
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1018..1407
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2
                     /note="binds to a specific stem-loop structure in the viral
                     RNA (Peabody, 1993)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     primer_bind     complement(1538..1555)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1538..1554)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1595..2182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2083..2102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2194..2221)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(2245..2358,1..702))
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
       61 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
      121 gacgactgaa tccggtgaga atggcaagag tttatgcatt tctttccaga cttgttcaac
      181 aggccagcca ttacgctcgt catcaaaatc actcgcacca accaaaccgt tattcattcg
      241 tgattgcgcc tgagcgagac gaaatacgcg atcgccgtta aaaggacaat tacaaacagg
      301 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
      361 aggatattct tctaatacct ggaatgctgt tttccctggg atcgcagtgg tgagtaacca
      421 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
      481 ccagtttagc ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
      541 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
      601 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
      661 tcgcggcttc aagcaagacg tttcccgttg aatatggctc attttagctt ccttagctcc
      721 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
      781 gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct tttgactttc
      841 tgctatggag gtcaggtatg atttaaatgg tcagtattga gcctcaggaa acagctatga
      901 catcaagctg actagataat ctagctgatc gtggaccgat catacgtata atgccgtaag
      961 atcacgcgcg aattcgaaga cgccatgccc aaaaagaaaa gaaaagtggg tagtatggcg
     1021 tctaacttca cccagttcgt tctggttgac aacggtggta ccggtgacgt taccgttgcg
     1081 ccgtctaact tcgcgaacgg tgttgcggaa tggatctctt ctaactctcg ttctcaggcg
     1141 tacaaagtta cctgctctgt tcgtcaatcc tccgcgcaga accgtaaata caccatcaaa
     1201 gttgaagttc cgaaagttgc gacccagacc gttggtggtg ttgaactgcc ggttgcggcg
     1261 tggcgttctt acctgaacat ggaactgacc atcccgatct tcgcgaccaa ctctgactgc
     1321 gaactgatcg ttaaagcgat gcagggtctg ctgaaagacg gtaacccgat cccgtctgcg
     1381 atcgcggcga actctggtat ctactaggct tgcgtcttcg aattcgaggc cgcatatcat
     1441 tcaggacgag cctcagactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     1501 cggtccacga tcagctagaa tcaagctgac tagataaact ggccgtcgtt ttacacgggt
     1561 gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1621 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1681 agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1741 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1801 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1861 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1921 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1981 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2041 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2101 ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt gatgctcgtc
     2161 aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc gaaggtgagc
     2221 caggtgatta catttaggtc ctcattagaa aaactcatcg agcatcaagt gaaactgcaa
     2281 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     2341 agaaaactca ccgaggca
//
