LOCUS       sequence-233427-        8509 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 8509)
  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 8509)
  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..8509
                     /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    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2622..2638
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2680..2713)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2784..3190
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3239..3248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3245..4180
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    join(4246..4820,4821..4823,4824..4832)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4413..4430)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4640..4659
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4833..5231
                     /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(5309..5342)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5398..5986
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5451..5471)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5869..5880)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5989..6005)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6515..6695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6757..7345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6837..6856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7516..8307,8308..8376))
                     /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     8139..8158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8377..8481)
                     /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 gccgcgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt tagtaccggg cccgctctag tcgaggtcga cggtatcgat
     2641 aagctcgctt cacgagattc cagcaggtcg agggacctaa taacttcgta tagcatacat
     2701 tatacgaagt tatattaagg gttccaagct taagcggccg gccgctgaaa gaccccacct
     2761 gtaggtttgg caagctagct gcagtaacgc cattttgcaa ggcatggaaa aataccaaac
     2821 caagaataga gaagttcaga tcaagggcgg gtacatgaaa atagctaacg ttgggccaaa
     2881 caggatatct gcggtgagca gtttcggccc cggcccgggg ccaagaacag atggtcaccg
     2941 cagtttcggc cccggcccga ggccaagaac agatggtccc cagatatggc ccaaccctca
     3001 gcagtttctt aagacccatc agatgtttcc aggctccccc aaggacctga aatgaccctg
     3061 cgccttattt gaattaacca atcagcctgc ttctcgcttc tgttcgcgcg cttctgcttc
     3121 ccgagctcta taaaagagct cacaacccct cactcggcgc gccagtcctc cgattgactg
     3181 agtcgcccgg atccaagctt gattcttctg acacaacagt ctcgaaccaa aggctggagc
     3241 caccatggct tccaaggtgt acgaccccga gcaacgcaaa cgcatgatca ctgggcctca
     3301 gtggtgggct cgctgcaagc aaatgaacgt gctggactcc ttcatcaact actatgattc
     3361 cgagaagcac gccgagaacg ccgtgatttt tctgcatggt aacgctgcct ccagctacct
     3421 gtggaggcac gtcgtgcctc acatcgagcc cgtggctaga tgcatcatcc ctgatctgat
     3481 cggaatgggt aagtccggca agagcgggaa tggctcatat cgcctcctgg atcactacaa
     3541 gtacctcacc gcttggttcg agctgctgaa ccttccaaag aaaatcatct ttgtgggcca
     3601 cgactggggg gcttgtctgg cctttcacta ctcctacgag caccaagaca agatcaaggc
     3661 catcgtccat gctgagagtg tcgtggacgt gatcgagtcc tgggacgagt ggcctgacat
     3721 cgaggaggat atcgccctga tcaagagcga agagggcgag aaaatggtgc ttgagaataa
     3781 cttcttcgtc gagaccatgc tcccaagcaa gatcatgcgg aaactggagc ctgaggagtt
     3841 cgctgcctac ctggagccat tcaaggagaa gggcgaggtt agacggccta ccctctcctg
     3901 gcctcgcgag atccctctcg ttaagggagg caagcccgac gtcgtccaga ttgtccgcaa
     3961 ctacaacgcc taccttcggg ccagcgacga tctgcctaag atgttcatcg agtccgaccc
     4021 tgggttcttt tccaacgcta ttgtcgaggg agctaagaag ttccctaaca ccgagttcgt
     4081 gaaggtgaag ggcctccact tcagccagga ggacgctcca gatgaaatgg gtaagtacat
     4141 caagagcttc gtggagcgcg tgctgaagaa cgagcagtaa gaattcctgc aggcctcgac
     4201 gagggccggc gcgccgcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc
     4261 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     4321 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     4381 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     4441 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     4501 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     4561 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     4621 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     4681 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     4741 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     4801 ttgaaaaaca cgatgataat atggccacaa ccatggccaa gcctttgtct caagaagaat
     4861 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     4921 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     4981 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     5041 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     5101 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     5161 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     5221 gggagggctg acaccggcgg catggacgag ctgtacaagt aaagcggccg gccgccagca
     5281 cagtggtcga aattcgtcga gggacctaat aacttcgtat agcatacatt atacgaagtt
     5341 atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct tatcgataat
     5401 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     5461 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5521 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5581 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5641 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5701 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5761 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     5821 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5881 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     5941 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
     6001 cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     6061 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     6121 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6181 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     6241 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg
     6301 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa
     6361 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg
     6421 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca
     6481 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg
     6541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6661 cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa
     6721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
