LOCUS       sequence-252005-        4627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lower Copy expression construct for MS2 phage like particles with 
            Esp3I cloning sites for packaged RNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4627)
  AUTHORS   de Martin Garrido N, Crone MA, Ramlaul K, Simpson PA, Freemont PS, 
            Aylett CHS
  TITLE     Bacteriophage MS2 displays unreported capsid variability assembling 
            T = 4 and mixed capsids.
  JOURNAL   Mol Microbiol. 2019 Oct 16. doi: 10.1111/mmi.14406.
  PUBMED    31618483
REFERENCE   2  (bases 1 to 4627)
  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: "Mol 
            Microbiol. 2019 Oct 16. doi: 10.1111/mmi.14406."
COMMENT     Sequence Label: pMC051-HisMS2_PLP_pac_LC
FEATURES             Location/Qualifiers
     source          1..4627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        314..332
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    333..357
                     /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        join(366..371,372..389,390..395)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             415..437
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1702..2088
                     /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"
     CDS             2131..2148
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2527..2574
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2620..2638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        2658..2676
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      2691..2738
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2836..2853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2946..3531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3026..3045)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3705..4499)
                     /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"
     promoter        complement(4500..4604)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg aggtaatacg actcactata ggggaattgt gagcggataa caattcccct
      361 ctagatttac actttatgct tccggctcgt atgtttctag agaattcaaa taattttgtt
      421 taactttaag aaggagattt aaattggcta tcgctgtagg tagccggaat tccattccta
      481 ggaggtttga cctgtgcgag cttttagtac ccttgatagg gagaacgaaa ccttcgtccc
      541 ctccgttcgc gtttacgcgg acggtgagac tgaagataac tcattctctt taaaatatcg
      601 ttcgaactgg actcccggtc gttttaactc gactggggcc aaaacgaaac agtggcacta
      661 cccctctccg tattcacggg gggcgttaag tgtcacatcg atagatcaag gtgcctacaa
      721 gcgaagtggg tcatcgtggg gtcgcccgta cgaggagaaa gccggtttcg gcttctccct
      781 cgacgcacgc tcctgctaca gcctcttccc tgtaagccaa aacttgactt acatcgaagt
      841 gccgcagaac gttgcgaacc gggcgtcgac cgaagtcctg caaaaggtca cccagggtaa
      901 ttttaacctt ggtgttgctt tagcagaggc caggtcgaca gcctcacaac tcgcgacgca
      961 aaccattgcg ctcgtgaagg cgtacactgc cgctcgtcgc ggtaattggc gccaggcgct
     1021 ccgctacctt gccctaaacg aagatcgaaa gtttcgatca aaacacgtgg ccggcagatg
     1081 gttggagttg cagttcggtt ggttaccact aatgagtgat atccagggtg catatgagat
     1141 gcttacgaag gttcaccttc aagagtttct tcctatgaga gccgtacgtc aggtcggtac
     1201 taacatcaag ttagatggcc gtctgtcgta tccagctgca aacttccaga caacgtgcaa
     1261 catatcgcga cgtatcgtga tatggtttta cataaacgat gcacgtttgg catggttgtc
     1321 gtccctaggt atcttgaacc cactaggtat agtgtgggaa aaggtgcctt tctcattcgt
     1381 tgtcgactgg ctcctacctg taggtaacat gctcgagggc cttacggccc ccgtgggatg
     1441 ctcctacatg tcaggaacag ttactgacgt aataacgggt gagtccatca taagcgttga
     1501 cgctccctac gggtggactg tggagagaca gggcactgct aaggcccaaa tctcagccat
     1561 gcatcgaggg gtacaatccg tatggccaac aactggcgcg tacgtaaagt ctcctttctc
     1621 gatggtccat accttagatg cgttagcatt aatcaggcaa cggctctcta gatagagccc
     1681 tcaaccggag tttgaagcat ggcttctaac tttactcagt tcgttctcgt cgacaatggc
     1741 ggaactggcg acgtgactgt cgccccaagc aacttcgcta acggggtcgc tgaatggatc
     1801 agctctaact cgcgttcaca ggcttacaaa gtaacctgta gcgttcgtca gagctctgcg
     1861 cagaatcgca aatacaccat caaagtcgag gtgcctaaag tggcaaccca gactgttggt
     1921 ggtgtagagc ttcctgtagc cgcatggcgt tcgtacttaa atatggaact aaccattcca
     1981 attttcgcta cgaattccga ctgcgagctt attgttaagg caatgcaagg tcttctaaaa
     2041 gatggaaacc cgattccctc agcaatcgca gcaaactccg gcatctacgc taactttact
     2101 cagttcgttc tcgtcgacaa tggcggtacc catcaccatc accatcacgg taccggcgac
     2161 gtgactgtcg ccccaagcaa cttcgctaac ggggtcgctg aatggatcag ctctaactcg
     2221 cgttcacagg cttacaaagt aacctgtagc gttcgtcaga gctctgcgca gaatcgcaaa
     2281 tacaccatca aagtcgaggt gcctaaagtg gcaacccaga ctgttggtgg tgtagagctt
     2341 cctgtagccg catggcgttc gtacttaaat atggaactaa ccattccaat tttcgctacg
     2401 aattccgact gcgagcttat tgttaaggca atgcaaggtc ttctaaaaga tggaaacccg
     2461 attccctcag caatcgcagc aaactccggc atctactaat agacgccggc caccgctgag
     2521 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtcgaac
     2581 agaaagtaat cgtattgtac acggccgcat aatcgaaatt aatacgactc actataggga
     2641 gacgaggtca cgtctcaaca tgaggatcac ccatgtccgc tgagcaataa ctagcataac
     2701 cccttggggc ctctaaacgg gtcttgaggg gttttttgta atttcactgc ccgctttcca
     2761 gtcgggaaac ctgtcgtgcc agctgcatta acatggtcat agctgtttcc ttgcgtattg
     2821 ggcgctctcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg taaagcctgg
     2881 ggtgcctaat gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2941 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3001 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3061 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3121 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3181 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3241 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3301 actggtaaca ggattagcag agcgaggtat gtaggcgcta cagagttctt gaagtggtgg
     3361 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttattagaa aaattcatcc
     3721 agcagacgat aaaacgcaat acgctggcta tccggtgccg caatgccata cagcaccaga
     3781 aaacgatccg cccattcgcc gcccagttct tccgcaatat cacgggtggc cagcgcaata
     3841 tcctgataac gatccgccac gcccagacgg ccgcaatcaa taaagccgct aaaacggcca
     3901 ttttccacca taatgttcgg caggcacgca tcaccatggg tcaccaccag atcttcgcca
     3961 tccggcatgc tcgctttcag acgcgcaaac agctctgccg gtgccaggcc ctgatgttct
     4021 tcatccagat catcctgatc caccaggccc gcttccatac gggtacgcgc acgttcaata
     4081 cgatgtttcg cctgatgatc aaacggacag gtcgccgggt ccagggtatg cagacgacgc
     4141 atggcatccg ccataatgct cactttttct gccggcgcca gatggctaga cagcagatcc
     4201 tgacccggca cttcgcccag cagcagccaa tcacggcccg cttcggtcac cacatccagc
     4261 accgccgcac acggaacacc ggtggtggcc agccagctca gacgcgccgc ttcatcctgc
     4321 agctcgttca gcgcaccgct cagatcggtt ttcacaaaca gcaccggacg accctgcgcg
     4381 ctcagacgaa acaccgccgc atcagagcag ccaatggtct gctgcgccca atcatagcca
     4441 aacagacgtt ccacccacgc tgccgggcta cccgcatgca ggccatcctg ttcaatcata
     4501 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4561 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4621 gtgccac
//
