LOCUS       sequence-202895-        6941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing  SNAP-KDEL for ER labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6941)
  AUTHORS   Voigt F, Zhang H, Cui XA, Triebold D, Liu AX, Eglinger J, Lee ES, 
            Chao JA, Palazzo AF
  TITLE     Single-Molecule Quantification of Translation-Dependent Association 
            of mRNAs with the Endoplasmic Reticulum.
  JOURNAL   Cell Rep. 2017 Dec 26;21(13):3740-3753. doi: 
            10.1016/j.celrep.2017.12.008.
  PUBMED    29281824
REFERENCE   2  (bases 1 to 6941)
  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.celrep.2017.12"; journalName: "Cell Rep"; date: 
            "2017-12-26- 26"; volume: "21"; issue: "13"; pages: "3740-3753"
COMMENT     Sequence Label: phage UbiC SNAP-KDEL
FEATURES             Location/Qualifiers
     source          1..6941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     sig_peptide     414..467
                     /codon_start=1
                     /product="signal sequence of chicken lysozyme"
                     /label=lysozyme signal sequence
                     /translation="MRSLLILVLCFLPLAALG"
     regulatory      516..525
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             522..1067
                     /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    1114..1702
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1167..1187)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1585..1596)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1777..2010
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2129..2147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2215..2319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2320..2388,2389..3180)
                     /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(2538..2557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3351..3939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3840..3859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4093..4110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4350..4371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4386..4391,4392..4409,4410..4416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4424..4440
                     /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     4429..4451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4448..4464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4448..4464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4514..4533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4725..5358
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5405..5530
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6027..6260
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6445..6489
                     /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    6782..6899
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg
       61 ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag
      121 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
      181 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
      241 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
      301 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
      361 cgcggttctt gtttgtggat cgctgtgatc gtcacttggc ggccgccgcc accatgaggt
      421 ctttgctaat cttggtgctt tgcttcctgc ccctggctgc tctggggaaa gtctttggac
      481 gatgtgagct ggcagcggct atgaagccac cggtcgccac catggacaaa gactgcgaaa
      541 tgaagcgcac caccctggat agccctctgg gcaagctgga actgtctggg tgcgaacagg
      601 gcctgcaccg tatcatcttc ctgggcaaag gaacatctgc cgccgacgcc gtggaagtgc
      661 ctgccccagc cgccgtgctg ggcggaccag agccactgat gcaggccacc gcctggctca
      721 acgcctactt tcaccagcct gaggccatcg aggagttccc tgtgccagcc ctgcaccacc
      781 cagtgttcca gcaggagagc tttacccgcc aggtgctgtg gaaactgctg aaagtggtga
      841 agttcggaga ggtcatcagc tacagccacc tggccgccct ggccggcaat cccgccgcca
      901 ccgccgccgt gaaaaccgcc ctgagcggaa atcccgtgcc cattctgatc ccctgccacc
      961 gggtggtgca gggcgacctg gacgtggggg gctacgaggg cgggctcgcc gtgaaagagt
     1021 ggctgctggc ccacgagggc cacagactgg gcaagcctgg gctgggtggg gggggggggt
     1081 ctaaggatga attgtaatga atcgatagat cctaatcaac ctctggatta caaaatttgt
     1141 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1201 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1261 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     1321 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     1381 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     1441 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     1501 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     1561 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     1621 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     1681 tccctttggg ccgcctcccc gcctgagatc ctttaagacc aatgacttac aaggcagctg
     1741 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     1801 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     1861 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     1921 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     1981 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
     2041 atttataact tgcaaagaaa tgaatatcag agagtgagag gcccgggtta attaaggaaa
     2101 gggctagatc attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2521 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3481 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3601 gataagtcgt gtcttactgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4141 cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga ctaatttttt
     4201 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtaaggag
     4261 gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag gtttcccgac
     4321 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     4381 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     4441 tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc atttttttca
     4501 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
     4561 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
     4621 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
     4681 taaagaaatt gtatttgtta aatatgtact acaaacttag tagttggaag ggctaattca
     4741 ctcccaaaga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     4801 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     4861 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     4921 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     4981 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     5041 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     5101 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     5161 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5221 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5281 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5341 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     5401 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     5461 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     5521 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     5581 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     5641 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     5701 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     5761 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     5821 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg
     5881 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     5941 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     6001 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     6061 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     6121 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     6181 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     6241 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     6301 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     6361 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     6421 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     6481 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     6541 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     6601 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     6661 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     6721 cgattagtga acggatctcg acggtatcgc cgaattcaca aatggcagta ttcatccaca
     6781 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     6841 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     6901 gggtttatta cagggacagc agagatccag tttggactag t
//
