LOCUS       sequence-227361-        8219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing G3BP1-SNAP fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8219)
  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 8219)
  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 G3BP1-SNAP
FEATURES             Location/Qualifiers
     source          1..8219
                     /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             4047..4589
                     /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="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     misc_feature    4609..5197
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4662..4682)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5080..5091)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5272..5505
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5624..5642)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5710..5814
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5815..5883,5884..6675)
                     /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(6033..6052)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6846..7434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7335..7354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7588..7605
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7845..7866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7881..7886,7887..7904,7905..7911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7919..7935
                     /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     7924..7946
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7943..7959
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7943..7959
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8009..8028
                     /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 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2521 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2581 ggttcttgtt tgtggatcgc tgtgatcgtc acttggcggc cgccgccacc atggtgatgg
     2641 agaagcctag tcccctgctg gtcgggcggg aatttgtgag acagtattac acactgctga
     2701 accaggcccc agacatgctg catagatttt atggaaagaa ctcttcttat gtccatgggg
     2761 gattggattc aaatggaaag ccagcagatg cagtctacgg acagaaagaa atccacagga
     2821 aagtgatgtc acaaaacttc accaactgcc acaccaagat tcgccatgtt gatgctcatg
     2881 ccacgctaaa tgatggtgtg gtagtccagg tgatggggct tctctctaac aacaaccagg
     2941 ctttgaggag attcatgcaa acgtttgtcc ttgctcctga ggggtctgtt gcaaataaat
     3001 tctatgttca caatgatatc ttcagatacc aagatgaggt ctttggtggg tttgtcactg
     3061 agcctcagga ggagtctgaa gaagaagtag aggaacctga agaaagacag caaacacctg
     3121 aggtggtacc tgatgattct ggaactttct atgatcaggc agttgtcagt aatgacatgg
     3181 aagaacattt agaggagcct gttgctgaac cagagcctga tcctgaacca gaaccagaac
     3241 aagaacctgt atctgaaatc caagaggaaa agcctgagcc agtattagaa gaaactgccc
     3301 ctgaggatgc tcagaagagt tcttctccag cacctgcaga catagctcag acagtacagg
     3361 aagacttgag gacattttct tgggcatctg tgaccagtaa gaatcttcca cccagtggag
     3421 ctgttccagt tactgggata ccacctcatg ttgttaaagt accagcttca cagccccgtc
     3481 cagagtctaa gcctgaatct cagattccac cacaaagacc tcagcgggat caaagagtgc
     3541 gagaacaacg aataaatatt cctccccaaa ggggacccag accaatccgt gaggctggtg
     3601 agcaaggtga cattgaaccc cgaagaatgg tgagacaccc tgacagtcac caactcttca
     3661 ttggcaacct gcctcatgaa gtggacaaat cagagcttaa agatttcttt caaagttatg
     3721 gaaacgtggt ggagttgcgc attaacagtg gtgggaaatt acccaatttt ggttttgttg
     3781 tgtttgatga ttctgagcct gttcagaaag tccttagcaa caggcccatc atgttcagag
     3841 gtgaggtccg tctgaatgtc gaagagaaga agactcgagc tgccagggaa ggcgaccgac
     3901 gagataatcg ccttcgggga cctggaggcc ctcgaggtgg gctgggtggt ggaatgagag
     3961 gccctccccg tggaggcatg gtgcagaaac caggatttgg agtgggaagg gggcttgcgc
     4021 cacggcaggg gggggggggg tctagagaca aagactgcga aatgaagcgc accaccctgg
     4081 atagccctct gggcaagctg gaactgtctg ggtgcgaaca gggcctgcac cgtatcatct
     4141 tcctgggcaa aggaacatct gccgccgacg ccgtggaagt gcctgcccca gccgccgtgc
     4201 tgggcggacc agagccactg atgcaggcca ccgcctggct caacgcctac tttcaccagc
     4261 ctgaggccat cgaggagttc cctgtgccag ccctgcacca cccagtgttc cagcaggaga
     4321 gctttacccg ccaggtgctg tggaaactgc tgaaagtggt gaagttcgga gaggtcatca
     4381 gctacagcca cctggccgcc ctggccggca atcccgccgc caccgccgcc gtgaaaaccg
     4441 ccctgagcgg aaatcccgtg cccattctga tcccctgcca ccgggtggtg cagggcgacc
     4501 tggacgtggg gggctacgag ggcgggctcg ccgtgaaaga gtggctgctg gcccacgagg
     4561 gccacagact gggcaagcct gggctgggtt aatgaatcga tagatcctaa tcaacctctg
     4621 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4681 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4741 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4801 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4861 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4921 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4981 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5041 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5101 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5161 cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta agaccaatga
     5221 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     5281 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     5341 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5401 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5461 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     5521 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcccg
     5581 ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt gatacgccta
     5641 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5761 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5821 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5881 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5941 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6001 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     6061 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6121 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6181 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6241 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6301 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6361 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6421 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6481 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6541 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6601 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6661 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6721 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6781 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6841 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6901 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6961 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7021 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7081 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7141 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7201 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7261 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7321 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7381 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7441 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7501 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7561 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7621 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag caagctcatg
     7681 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     7741 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc cccgtggcac
     7801 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7861 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7921 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttcacaaat
     7981 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     8041 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     8101 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     8161 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
