LOCUS       sequence-234203-        8456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing tagRFP-T-DDX6 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8456)
  AUTHORS   Wilbertz JH, Voigt F, Horvathova I, Roth G, Zhan Y, Chao JA
  TITLE     Single-Molecule Imaging of mRNA Localization and Regulation during 
            the Integrated Stress Response.
  JOURNAL   Mol Cell. 2019 Jan 4. pii: S1097-2765(18)31036-0. doi: 
            10.1016/j.molcel.2018.12.006.
  PUBMED    30661979
REFERENCE   2  (bases 1 to 8456)
  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 Cell. 
            2019 Jan 4. pii: S1097-2765(18)31036-0. doi: 
            10.1016/j.molcel.2018.12.006."
COMMENT     Sequence Label: phage UbiC tagRFP-T-DDX6
FEATURES             Location/Qualifiers
     source          1..8456
                     /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      2625..2634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4846..5434
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4899..4919)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5317..5328)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5509..5742
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5861..5879)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5947..6051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6052..6120,6121..6912)
                     /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(6270..6289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7083..7671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7572..7591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7825..7842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8082..8103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8118..8123,8124..8141,8142..8148)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8156..8172
                     /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     8161..8183
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8180..8196
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8180..8196
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8246..8265
                     /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 aatcttgcct
      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 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgccgccacc atggtgagca
     2641 agggcgagga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga
     2701 acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga
     2761 ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta
     2821 ccagcttcat gtacggcagc agaaccttca tcaaccacac ccagggcatc cccgacttct
     2881 ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg
     2941 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca
     3001 agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct
     3061 gggaggccaa caccgagatg ctgtaccccg ctgacggcgg cctggaaggc agaaccgaca
     3121 tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttcaagacc acatacagat
     3181 ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac cacagactgg
     3241 aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca
     3301 gatactgcga cctccctagc aaactggggc acaaacttaa tggcatggac gagctgtaca
     3361 aggggggggg ggggtctaga agcacggcca gaacagagaa ccctgttata atgggtctgt
     3421 ccagtcaaaa tggtcagctg agaggccctg tgaaacccac tggtggccct ggaggagggg
     3481 gcacacagac acagcaacag atgaaccagc tgaaaaacac caacacaatc aataatggca
     3541 ctcagcagca agcacagagt atgaccacca ctattaaacc tggtgatgac tggaaaaaga
     3601 ctttaaaact ccctccaaag gatctaagaa tcaaaacttc ggatgtgacc tccacaaaag
     3661 gaaatgagtt tgaagattac tgtttgaaac gggagttact gatgggaatt tttgaaatgg
     3721 gctgggaaaa gccatctcct attcaggagg agagcattcc cattgcttta tctggtaggg
     3781 atatcttagc tagagcaaaa aatggaacag gcaagagcgg tgcctacctc attcccttac
     3841 ttgaacggct agacctgaag aaggacaata tacaagcaat ggtgattgtt cccactagag
     3901 aacttgctct acaggtcagt caaatttgca tccaggtcag caaacacatg ggaggggcca
     3961 aagtgatggc aaccacagga ggaaccaatt tacgagatga cataatgagg cttgatgata
     4021 cagtgcacgt ggtgattgct acccctggga gaatcctgga tcttattaag aaaggagtag
     4081 caaaggttga tcatgtccag atgatagtat tggatgaggc agataagttg ctgtcacagg
     4141 attttgtgca gataatggag gatattattc tcacgctacc taaaaacagg cagattttac
     4201 tatattccgc tactttccct cttagtgtac agaagttcat gaattcccat ttgcagaaac
     4261 cctatgagat taacctgatg gaggaactaa ctctgaaggg agtaacccag tactacgcat
     4321 atgtaactga gcgccaaaaa gtacactgcc tcaacacact tttctccagg cttcagataa
     4381 accagtcgat cattttctgt aactcctctc agcgagttga attgctagcc aagaagattt
     4441 ctcaactggg ttattcttgc ttctatattc atgctaaaat gaggcaggaa catcgaaatc
     4501 gtgtatttca tgatttccga aatggcttat gccgcaatct tgtttgcact gatctgttta
     4561 cccgaggtat tgatatacaa gctgtgaatg tggtaataaa ctttgatttc ccaaagctgg
     4621 cagagaccta tctccatcgt attggaagat caggtcgctt tggtcatctt ggcttagcca
     4681 tcaacttgat cacatatgat gatcgcttca acctgaaaag tattgaggag cagctgggaa
     4741 cagaaattaa acctattccg agcaacattg ataagagcct gtatgtggca gaataccaca
     4801 gcgagcctgt agaagatgag aaaccttaat gaatcgatag atcctaatca acctctggat
     4861 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     4921 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     4981 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5041 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5101 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5161 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5221 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5281 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5341 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     5401 acgagtcgga tctccctttg ggccgcctcc ccgcctgaga tcctttaaga ccaatgactt
     5461 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     5521 ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc
     5581 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     5641 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     5701 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc
     5761 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggcccgggt
     5821 taattaagga aagggctaga tcattcttga agacgaaagg gcctcgtgat acgcctattt
     5881 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5941 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6001 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6061 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6121 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6181 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6241 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     6301 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6361 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6421 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6481 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6541 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6601 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6661 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6721 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6781 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6841 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6901 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6961 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7021 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7081 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7141 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7201 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7261 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7321 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7381 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7441 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7501 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7561 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7621 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7681 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7741 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7801 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7861 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct
     7921 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     7981 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccc gtggcacgac
     8041 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8101 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8161 agcggataac aatttcacac aggaaacagc tatgacatga ttacgaattt cacaaataaa
     8221 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     8281 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
     8341 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
     8401 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
