LOCUS       sequence-237571-        8270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing SNAP-DDX6 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8270)
  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 8270)
  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 SNAP-DDX6
FEATURES             Location/Qualifiers
     source          1..8270
                     /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"
     CDS             2631..3176
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    4660..5248
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4713..4733)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5131..5142)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5323..5556
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5675..5693)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5761..5865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5866..5934,5935..6726)
                     /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(6084..6103)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6897..7485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7386..7405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7639..7656
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7896..7917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7932..7937,7938..7955,7956..7962)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7970..7986
                     /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     7975..7997
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7994..8010
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7994..8010
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8060..8079
                     /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 atggacaaag
     2641 actgcgaaat gaagcgcacc accctggata gccctctggg caagctggaa ctgtctgggt
     2701 gcgaacaggg cctgcaccgt atcatcttcc tgggcaaagg aacatctgcc gccgacgccg
     2761 tggaagtgcc tgccccagcc gccgtgctgg gcggaccaga gccactgatg caggccaccg
     2821 cctggctcaa cgcctacttt caccagcctg aggccatcga ggagttccct gtgccagccc
     2881 tgcaccaccc agtgttccag caggagagct ttacccgcca ggtgctgtgg aaactgctga
     2941 aagtggtgaa gttcggagag gtcatcagct acagccacct ggccgccctg gccggcaatc
     3001 ccgccgccac cgccgccgtg aaaaccgccc tgagcggaaa tcccgtgccc attctgatcc
     3061 cctgccaccg ggtggtgcag ggcgacctgg acgtgggggg ctacgagggc gggctcgccg
     3121 tgaaagagtg gctgctggcc cacgagggcc acagactggg caagcctggg ctgggtgggg
     3181 ggggggggtc tagaagcacg gccagaacag agaaccctgt tataatgggt ctgtccagtc
     3241 aaaatggtca gctgagaggc cctgtgaaac ccactggtgg ccctggagga gggggcacac
     3301 agacacagca acagatgaac cagctgaaaa acaccaacac aatcaataat ggcactcagc
     3361 agcaagcaca gagtatgacc accactatta aacctggtga tgactggaaa aagactttaa
     3421 aactccctcc aaaggatcta agaatcaaaa cttcggatgt gacctccaca aaaggaaatg
     3481 agtttgaaga ttactgtttg aaacgggagt tactgatggg aatttttgaa atgggctggg
     3541 aaaagccatc tcctattcag gaggagagca ttcccattgc tttatctggt agggatatct
     3601 tagctagagc aaaaaatgga acaggcaaga gcggtgccta cctcattccc ttacttgaac
     3661 ggctagacct gaagaaggac aatatacaag caatggtgat tgttcccact agagaacttg
     3721 ctctacaggt cagtcaaatt tgcatccagg tcagcaaaca catgggaggg gccaaagtga
     3781 tggcaaccac aggaggaacc aatttacgag atgacataat gaggcttgat gatacagtgc
     3841 acgtggtgat tgctacccct gggagaatcc tggatcttat taagaaagga gtagcaaagg
     3901 ttgatcatgt ccagatgata gtattggatg aggcagataa gttgctgtca caggattttg
     3961 tgcagataat ggaggatatt attctcacgc tacctaaaaa caggcagatt ttactatatt
     4021 ccgctacttt ccctcttagt gtacagaagt tcatgaattc ccatttgcag aaaccctatg
     4081 agattaacct gatggaggaa ctaactctga agggagtaac ccagtactac gcatatgtaa
     4141 ctgagcgcca aaaagtacac tgcctcaaca cacttttctc caggcttcag ataaaccagt
     4201 cgatcatttt ctgtaactcc tctcagcgag ttgaattgct agccaagaag atttctcaac
     4261 tgggttattc ttgcttctat attcatgcta aaatgaggca ggaacatcga aatcgtgtat
     4321 ttcatgattt ccgaaatggc ttatgccgca atcttgtttg cactgatctg tttacccgag
     4381 gtattgatat acaagctgtg aatgtggtaa taaactttga tttcccaaag ctggcagaga
     4441 cctatctcca tcgtattgga agatcaggtc gctttggtca tcttggctta gccatcaact
     4501 tgatcacata tgatgatcgc ttcaacctga aaagtattga ggagcagctg ggaacagaaa
     4561 ttaaacctat tccgagcaac attgataaga gcctgtatgt ggcagaatac cacagcgagc
     4621 ctgtagaaga tgagaaacct taatgaatcg atagatccta atcaacctct ggattacaaa
     4681 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4741 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4801 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4861 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4921 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4981 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     5041 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     5101 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     5161 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     5221 cggatctccc tttgggccgc ctccccgcct gagatccttt aagaccaatg acttacaagg
     5281 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5341 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5401 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5461 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5521 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     5581 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggccc gggttaatta
     5641 aggaaagggc tagatcattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     5701 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5761 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5821 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5881 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5941 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6001 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6061 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     6121 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6181 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6241 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6301 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6361 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6421 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6481 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6541 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6601 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6661 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6721 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6781 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6841 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6901 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6961 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7021 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7081 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7141 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7201 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7261 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7321 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7381 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7441 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7501 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7561 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     7621 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     7681 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gcaagctcat ggctgactaa
     7741 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     7801 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ccccgtggca cgacaggttt
     7861 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7921 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7981 taacaatttc acacaggaaa cagctatgac atgattacga atttcacaaa taaagcattt
     8041 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     8101 atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt
     8161 accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt
     8221 tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
