LOCUS       sequence-237229-        8506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for stable expression of Renilla luciferase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8506)
  AUTHORS   Gorbea C, Mosbruger T, Nix DA, Cazalla D
  TITLE     Viral miRNA adaptor differentially recruits miRNAs to target mRNAs 
            through alternative base-pairing.
  JOURNAL   Elife. 2019 Sep 20;8. pii: 50530. doi: 10.7554/eLife.50530.
  PUBMED    31538617
REFERENCE   2  (bases 1 to 8506)
  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: "Elife."; 
            date: "2019-09-20"; volume: "8. pii: 50530. doi"; pages: " 
            10.7554/eLife.50530"
COMMENT     Sequence Label: LeGo-Renilla-BLAST
FEATURES             Location/Qualifiers
     source          1..8506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2619..2635
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2677..2710)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2781..3187
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3236..3245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3242..4177
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    join(4243..4817,4818..4820,4821..4829)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4410..4427)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4637..4656
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4830..5228
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(5306..5339)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5395..5983
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5448..5468)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5866..5877)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5986..6002)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6512..6692
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6754..7342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6834..6853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7513..8304,8305..8373))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8136..8155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8374..8478)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgctctatcg aggtcgacgg tatcgataag
     2641 ctcgcttcac gagattccag caggtcgagg gacctaataa cttcgtatag catacattat
     2701 acgaagttat attaagggtt ccaagcttaa gcggccggcc gctgaaagac cccacctgta
     2761 ggtttggcaa gctagctgca gtaacgccat tttgcaaggc atggaaaaat accaaaccaa
     2821 gaatagagaa gttcagatca agggcgggta catgaaaata gctaacgttg ggccaaacag
     2881 gatatctgcg gtgagcagtt tcggccccgg cccggggcca agaacagatg gtcaccgcag
     2941 tttcggcccc ggcccgaggc caagaacaga tggtccccag atatggccca accctcagca
     3001 gtttcttaag acccatcaga tgtttccagg ctcccccaag gacctgaaat gaccctgcgc
     3061 cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg
     3121 agctctataa aagagctcac aacccctcac tcggcgcgcc agtcctccga ttgactgagt
     3181 cgcccggatc caagcttgat tcttctgaca caacagtctc gaaccaaagg ctggagccac
     3241 catggcttcc aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg
     3301 gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga
     3361 gaagcacgcc gagaacgccg tgatttttct gcatggtaac gctgcctcca gctacctgtg
     3421 gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg
     3481 aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta
     3541 cctcaccgct tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga
     3601 ctggggggct tgtctggcct ttcactactc ctacgagcac caagacaaga tcaaggccat
     3661 cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga
     3721 ggaggatatc gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt
     3781 cttcgtcgag accatgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc
     3841 tgcctacctg gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc
     3901 tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta
     3961 caacgcctac cttcgggcca gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg
     4021 gttcttttcc aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa
     4081 ggtgaagggc ctccacttca gccaggagga cgctccagat gaaatgggta agtacatcaa
     4141 gagcttcgtg gagcgcgtgc tgaagaacga gcagtaagaa ttcctgcagg cctcgacgag
     4201 ggccggcgcg ccgcggccgc tacgtaaatt ccgccccccc cccccctctc cctccccccc
     4261 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     4321 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     4381 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     4441 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     4501 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     4561 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     4621 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     4681 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     4741 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     4801 aaaaacacga tgataatatg gccacaacca tggccaagcc tttgtctcaa gaagaatcca
     4861 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     4921 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     4981 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     5041 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     5101 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     5161 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     5221 agggctgaca ccggcggcat ggacgagctg tacaagtaaa gcggccggcc gccagcacag
     5281 tggtcgaaat tcgtcgaggg acctaataac ttcgtatagc atacattata cgaagttata
     5341 catgtttaag ggttccggtt ccactaggta caattcgata tcaagcttat cgataatcaa
     5401 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5461 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5521 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     5581 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     5641 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     5701 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     5761 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     5821 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5881 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5941 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     6001 cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     6061 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     6121 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     6181 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
     6241 gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat
     6301 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt
     6361 agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat
     6421 ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca
     6481 catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc
     6541 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6601 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6661 acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc
     6721 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6781 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6841 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6901 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6961 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7021 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7081 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7141 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7201 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7261 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7321 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7381 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7441 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7501 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7561 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7621 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7681 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7741 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7801 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7861 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7921 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7981 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8041 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8101 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8161 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8221 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8281 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8341 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8401 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8461 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
