LOCUS       sequence-325003-        8317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing synonymous tandem MS2 coat protein 
            fused to synonymous tandem Halo-tag and the RH1 domain of RILPL2. 
            Used for single-molecule imaging of actin-anchored mRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8317)
  AUTHORS   Mateju D, Eichenberger B, Voigt F, Eglinger J, Roth G, Chao JA
  TITLE     Single-Molecule Imaging Reveals Translation of mRNAs Localized to 
            Stress Granules.
  JOURNAL   Cell. 2020 Dec 23;183(7):1801-1812.e13. doi: 
            10.1016/j.cell.2020.11.010. Epub 2020 Dec 11.
  PUBMED    33308477
REFERENCE   2  (bases 1 to 8317)
  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: 
            "10.1016/j.cell.2020.11.010"; journalName: "Cell"; date: 
            "2020-12-23- 23"; volume: "183"; issue: "7"; pages: "1801-1812.e13"
COMMENT     Sequence Label: pHAGE UbiC NLS-HA-stdMCP-stdHalo-RH1
FEATURES             Location/Qualifiers
     source          1..8317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2218..2616
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      2624..2633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2636..2656
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2660..2686
                     /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             2660..2686
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2687..2710
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     regulatory      3443..3452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3449..4336
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     CDS             4349..4372
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    4707..5295
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4760..4780)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5178..5189)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5370..5603
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5722..5740)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5808..5912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5913..5981,5982..6773)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6131..6150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6944..7532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7433..7452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7686..7703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7943..7964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7979..7984,7985..8002,8003..8009)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8017..8033
                     /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)."
     primer_bind     8022..8044
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8041..8057
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8041..8057
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8107..8126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtggc
     2221 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
     2281 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2341 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2401 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     2461 agtagtccct tctcggcgat tctgtggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgcgccacca tgggcccaaa
     2641 aaagaaaaga aaagttggct acccctacga cgtgcccgac tacgccctgg aagttctgtt
     2701 ccaggggccc ctgttattaa ttaacgcttc taactttact cagttcgttc tcgtcgacaa
     2761 tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc gctaacggga tcgctgaatg
     2821 gatcagctct aactcgcgtt cacaggctta caaagtaacc tgtagcgttc gtcagagctc
     2881 tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct aaaggcgcct ggcgttcgta
     2941 cttaaatatg gaactaacca ttccaatttt cgccacgaat tccgactgcg agcttattgt
     3001 taaggcaatg caaggtctcc taaaagatgg aaacccgatt ccctcagcaa tcgcagcaaa
     3061 ctccggcatc tacgccatgg ccagcaactt cacccagttc gtgctggtgg acaacggcgg
     3121 caccggcgac gtgaccgtgg cccccagcaa cttcgccaac ggcatcgccg agtggatcag
     3181 cagcaacagc agaagccagg cctacaaggt gacctgcagc gtgagacaga gcagcgccca
     3241 gaacagaaag tacaccatca aggtggaggt gcccaagggc gcctggagaa gctacctgaa
     3301 catggagctg accatcccca tcttcgccac caacagcgac tgcgagctga tcgtgaaggc
     3361 catgcagggc ctgctgaagg acggcaaccc catccccagc gccatcgccg ccaacagcgg
     3421 catctacgcc gacccaccgg tcgccaccat ggagatcggc accggcttcc cctttgaccc
     3481 tcactacgtg gaggtgctgg gcgagaggat gcactatgtg gacgtgggac caagagatgg
     3541 aacccccgtg ctgttcctgc acggcaaccc cacaagctcc tacgtgtggc ggaatatcat
     3601 cccacacgtg gcccccacac acagatgcat cgccccagat ctgatcggca tgggcaagtc
     3661 cgacaagccc gatctgggct atttctttga cgatcacgtg cggttcatgg acgcctttat
     3721 cgaggccctg ggcctggagg aggtggtgct ggtcatccac gattggggct ctgccctggg
     3781 ctttcactgg gccaagcgga accccgagag agtgaagggc atcgccttca tggagtttat
     3841 cagaccaatc ccaacctggg atgagtggcc tgagttcgcc agagagacat tccaggcctt
     3901 tcggaccaca gacgtgggca gaaagctgat catcgatcag aacgtgttta tcgaaggcac
     3961 cctgcctatg ggagtggtga ggccactgac agaggtggag atggaccact accgcgagcc
     4021 tttcctgaat ccagtggata gggagcctct gtggcgcttt cccaacgagc tgcctatcgc
     4081 cggcgagcca gccaatatcg tggccctggt ggaggagtat atggactggc tgcaccagtc
     4141 ccctgtgcca aagctgctgt tctggggcac cccaggcgtg ctgatccctc ccgcagaggc
     4201 agcaaggctg gccaagtctc tgcccaattg taaggccgtg gatatcggcc ctggcctgaa
     4261 cctgctgcag gaggacaatc ctgatctgat cggcagcgag atcgcccgct ggctgtctac
     4321 actggagatc agcggcggga gtggcggtct ggaagttctg tttcagggcc cgatggagga
     4381 gccccctgtg cgagaagagg aagaggagga gggagaggag gacgaggaga gggacgaggt
     4441 tgggcccgag ggggcgctgg gcaagagccc cttccagctg accgccgagg acgtgtatga
     4501 catctcctac ctgttgggcc gcgagcttat ggccctgggc agcgaccccc gggtgacgca
     4561 gctgcagttc aaagtcgtcc gcgtcctgga gatgctggag gcgctggtga atgagggcag
     4621 cctggcgctg gaggagctga agatggagag ggaccacctc aggaaggagg tggaggggct
     4681 gcggagacag taaatcgata gatcctaatc aacctctgga ttacaaaatt tgtgaaagat
     4741 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4801 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4861 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     4921 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     4981 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5041 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5101 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5161 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5221 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5281 gggccgcctc cccgcctgag atcctttaag accaatgact tacaaggcag ctgtagatct
     5341 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     5401 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     5461 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     5521 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     5581 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     5641 acttgcaaag aaatgaatat cagagagtga gaggcccggg ttaattaagg aaagggctag
     5701 atcattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5761 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5821 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5881 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5941 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6001 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6061 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6121 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     6181 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6241 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6301 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6361 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6421 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6481 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6541 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6601 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6661 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6721 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6781 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6841 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6901 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6961 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7021 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7081 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7141 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7201 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7261 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7321 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7381 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7441 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7501 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7561 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7621 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7681 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7741 ccgcgcgttg gccgattcat taatgcagca agctcatggc tgactaattt tttttattta
     7801 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     7861 tggaggccta ggcttttgca aaaagctccc cgtggcacga caggtttccc gactggaaag
     7921 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     7981 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     8041 caggaaacag ctatgacatg attacgaatt tcacaaataa agcatttttt tcactgcatt
     8101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aactggataa
     8161 ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc actgccaatt
     8221 acctgtggtt tcatttactc taaacctgtg attcctctga attattttca ttttaaagaa
     8281 attgtatttg ttaaatatgt actacaaact tagtagt
//
