LOCUS       sequence-175252-        6361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector expressing mutant version of the 
            reporter gene-Renilla Luciferase (Rluc8), which is more stable in 
            mouse serum and has more light output..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6361)
  AUTHORS   Loening AM, Fenn TD, Wu AM, Gambhir SS
  TITLE     Consensus guided mutagenesis of Renilla luciferase yields enhanced 
            stability and light output.
  JOURNAL   Protein Eng Des Sel. 2006 Sep;19(9):391-400. Epub 2006 Jul 20.
  PUBMED    16857694
REFERENCE   2  (bases 1 to 6361)
  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: "Protein Eng
            Des Sel."; date: "2006-09"; volume: "19(9)"; pages: "391-400. Epub 
            2006 Jul 20"
COMMENT     Sequence Label: pcDNA-RLuc8
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..936
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGAALAFHYAYEHQDRIKAIVHMESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETVLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
     primer_bind     complement(1048..1065)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1054..1278
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1324..1752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1411..1430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1621..1642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1761..1781)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1766..2095
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1946..2081
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2008..2027
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2162..2956
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2216..2235)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2826..2845
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3130..3251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3167..3186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3221..3240
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3300..3316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3300..3316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3313..3335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3324..3340
                     /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)."
     promoter        complement(join(3348..3354,3355..3372,3373..3378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3393..3414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3531..3548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3702..4290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3782..3801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4461..5252,5253..5321))
                     /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     5084..5103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5322..5426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5501..5520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5692..6071
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6072..6275
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6226..6246
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6320..6339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6320..6338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(6356..6361,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggcttcca aggtgtacga ccccgagcaa cgcaaacgca tgatcactgg gcctcagtgg
       61 tgggctcgct gcaagcaaat gaacgtgctg gactccttca tcaactacta tgattccgag
      121 aagcacgccg agaacgccgt gatttttctg catggtaacg ctacctccag ctacctgtgg
      181 aggcacgtcg tgcctcacat cgagcccgtg gctagatgca tcatccctga tctgatcgga
      241 atgggtaagt ccggcaagag cgggaatggc tcatatcgcc tcctggatca ctacaagtac
      301 ctcaccgctt ggttcgagct gctgaacctt ccaaagaaaa tcatctttgt gggccacgac
      361 tggggggctg ctctggcctt tcactacgcc tacgagcacc aagacaggat caaggccatc
      421 gtccatatgg agagtgtcgt ggacgtgatc gagtcctggg acgagtggcc tgacatcgag
      481 gaggatatcg ccctgatcaa gagcgaagag ggcgagaaaa tggtgcttga gaataacttc
      541 ttcgtcgaga ccgtgctccc aagcaagatc atgcggaaac tggagcctga ggagttcgct
      601 gcctacctgg agccattcaa ggagaagggc gaggttagac ggcctaccct ctcctggcct
      661 cgcgagatcc ctctcgttaa gggaggcaag cccgacgtcg tccagattgt ccgcaactac
      721 aacgcctacc ttcgggccag cgacgatctg cctaagctgt tcatcgagtc cgaccctggg
      781 ttcttttcca acgctattgt cgagggagct aagaagttcc ctaacaccga gttcgtgaag
      841 gtgaagggcc tccacttcct ccaggaggac gctccagatg aaatgggtaa gtacatcaag
      901 agcttcgtgg agcgcgtgct gaagaacgag cagtaaaagc ttggtaccga gctcggatcc
      961 actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta
     1021 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     1081 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1141 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1201 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     1261 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     1321 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1381 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1441 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1501 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1561 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1621 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1681 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1741 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     1801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     1861 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1921 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     1981 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     2041 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2101 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     2161 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2221 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2281 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2341 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2401 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2461 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2521 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2581 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2641 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     2701 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     2761 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     2821 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     2881 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     2941 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3001 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3061 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3121 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3181 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3241 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4141 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4441 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4501 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4561 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4621 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4681 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4741 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4801 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4861 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4921 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4981 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5041 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5101 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5161 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5221 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5281 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5341 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5401 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     5461 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     5521 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     5581 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     5641 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     5701 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5761 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5821 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     5881 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     5941 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6001 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6061 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     6121 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6181 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     6241 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     6301 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagccac
     6361 c
//
