LOCUS       sequence-323591-        7562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MS2 coat protein fused to GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7562)
  AUTHORS   Coulon A, Ferguson ML, de Turris V, Palangat M, Chow CC, Larson DR
  TITLE     Kinetic competition during the transcription cycle results in 
            stochastic RNA processing.
  JOURNAL   Elife. 2014 Oct 1;3. doi: 10.7554/eLife.03939.
  PUBMED    25271374
REFERENCE   2  (bases 1 to 7562)
  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: "Elife."; 
            date: "2014-10-1"; volume: "3. doi"; pages: " 10.7554/eLife.03939"
COMMENT     Sequence Label: MS2_GFP
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    254..278
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             287..307
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             311..337
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             311..337
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             338..349
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(726..750)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(752..754,755..1468)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(752..754,755..1465)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(794..815)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1055..1074)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1402..1423
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1470..1490)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     1558..1574
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1649..2237
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1702..1722)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2120..2131)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2240..2256)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             2766..2946
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3008..3596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3088..3107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3767..4558,4559..4627))
                     /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     4390..4409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4628..4732)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4807..4826)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4998..5377
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5379..5577
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5532..5552
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5595..5775
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5822..5947
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6444..6677
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6862..6906
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7204..7321
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    7389..7409
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
       61 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
      121 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
      181 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
      241 gtgatcgtca cttacaactt tgtacaaaaa agttggcacc atgggcccaa aaaagaaaag
      301 aaaagttggc tacccctacg acgtgcccga ctacgccatc gaaggccgcc atatgctagc
      361 cgttaaaatg gcttctaact ttactcagtt cgttctcgtc gacaatggcg gaactggcga
      421 cgtgactgtc gccccaagca acttcgctaa cgggatcgct gaatggatca gctctaactc
      481 gcgttcacag gcttacaaag taacctgtag cgttcgtcag agctctgcgc agaatcgcaa
      541 atacaccatc aaagtcgagg tgcctaaagg cgcctggcgt tcgtacttaa atatggaact
      601 aaccattcca attttcgcca cgaattccga ctgcgagctt attgttaagg caatgcaagg
      661 tctcctaaaa gatggaaacc cgattccctc agcaatcgca gcaaactccg gcatctacgc
      721 ggattcttca actttcttgt acaaagtggg cgtgagcaag ggcgaggagc tgttcaccgg
      781 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      841 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
      901 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
      961 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1021 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1081 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1141 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1201 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     1261 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     1321 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     1381 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1441 tctcggcatg gacgagctgt acaagtaatc aactttatta tacatagttg ataattcaag
     1501 ggatctaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatcga
     1561 ggtcgacggt atcgataagc tcgcttcacg agatcatgtt taagggttcc ggttccacta
     1621 ggtacaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     1681 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1741 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1801 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     1861 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     1921 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     1981 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2041 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2101 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2161 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2221 ttgggccgcc tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag
     2281 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     2341 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     2401 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     2461 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     2521 ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt
     2581 gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca
     2641 cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag
     2701 agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact
     2761 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     2821 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     2881 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     2941 ctagcagcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3001 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3061 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3121 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3181 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3241 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3301 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3361 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3421 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3481 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3541 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3601 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3661 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3721 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3781 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3841 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3901 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3961 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4021 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4081 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4141 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4201 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4261 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4321 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4381 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4441 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4501 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4561 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4621 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4681 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4741 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     4801 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     4861 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     4921 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     4981 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     5041 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     5101 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     5161 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     5221 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     5281 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     5341 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     5401 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     5461 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     5521 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagcgc
     5581 gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5641 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5701 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5761 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     5821 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     5881 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     5941 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     6001 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     6061 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     6121 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     6181 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     6241 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc
     6301 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     6361 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     6421 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     6481 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     6541 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     6601 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     6661 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     6721 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     6781 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     6841 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     6901 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     6961 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     7021 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     7081 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     7141 ttagtgaacg gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca
     7201 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     7261 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     7321 tcgggtttat tacagggaca gcagagatcc agtttggtta gtaccgggcc cgctctagat
     7381 tgaattatca actttgtata gaaaagttgg cgactagtgg cctccgcgcc gggttttggc
     7441 gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag
     7501 cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     7561 gg
//
