LOCUS       sequence-405438-        8122 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of Rluc-6xbsMS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8122)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 8122)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pcDNA3.1-beta-hRLuc-V5-6xbsMS2_V
FEATURES             Location/Qualifiers
     source          1..8122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2133..3065
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             3066..3107
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_RNA        3207..3225
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3246..3264
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3285..3303
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3324..3342
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3363..3381
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        3402..3420
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        complement(3442..3460)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3443..3460)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3480..3497)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3486..3710
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3756..4184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3843..3862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4053..4074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4193..4213)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4198..4527
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4378..4513
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4440..4459
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4594..5388
                     /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(4648..4667)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5258..5277
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5562..5683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5599..5618)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5653..5672
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5732..5748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5732..5748)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5745..5767)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5756..5772
                     /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(5780..5786,5787..5804,5805..5810))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5825..5846
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5963..5980)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6134..6721)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6214..6233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6892..7683,7684..7752))
                     /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     7515..7534
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7753..7857)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7932..7951)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 acatttgctt ctgacacaac tgtgttcact agcaacctca aacagacacc atggtgcacc
      721 tgactcctga ggagaagtct gccgttactg ccctgtgggg caaggtgaac gtggatgaag
      781 ttggtggtga ggccctgggc aggttggtat caaggttaca agacaggttt aaggagacca
      841 atagaaactg ggcatgtgga gacagagaag actcttgggt ttctgatagg cactgactct
      901 ctctgcctat tggtctattt tcccaccctt aggctgctgg tggtctaccc ttggacccag
      961 aggttctttg agtcctttgg ggatctgtcc actcctgatg ctgttatggg caaccctaag
     1021 gtgaaggctc atggcaagaa agtgctcggt gcctttagtg atggcctggc tcacctggac
     1081 aacctcaagg gcacctttgc cacactgagt gagctgcact gtgacaagct gcacgtggat
     1141 cctgagaact tcagggtgag tctatgggac ccttgatgtt ttctttcccc ttcttttcta
     1201 tggttaagtt catgtcatag gaaggggata agtaacaggg tacagtttag aatgggaaac
     1261 agacgaatga ttgcatcagt gtggaagtct caggatcgtt ttagtttctt ttatttgctg
     1321 ttcataacaa ttgttttctt ttgtttaatt cttgctttct ttttttttct tctccgcaat
     1381 ttttactatt atacttaatg ccttaacatt gtgtataaca aaaggaaata tctctgagat
     1441 acattaagta acttaaaaaa aaaactttac acagtctgcc tagtacatta ctatttggaa
     1501 tatatgtgtg cttatttgca tattcataat ctccctactt tattttcttt tatttttaat
     1561 tgatacataa tcattataca tatttatggg ttaaagtgta atgttttaat atgtgtacac
     1621 atattgacca aatcagggta attttgcatt tgtaatttta aaaaatgctt tcttctttta
     1681 atatactttt ttgtttatct tatttctaat actttcccta atctctttct ttcagggcaa
     1741 taatgataca atgtatcatg cctctttgca ccattctaaa gaataacagt gataatttct
     1801 gggttaaggc aatagcaata tttctgcata taaatatttc tgcatataaa ttgtaactga
     1861 tgtaagaggt ttcatattgc taatagcagc tacaatccag ctaccattct gcttttattt
     1921 tatggttggg ataaggctgg attattctga gtccaagcta ggcccttttg ctaatcatgt
     1981 tcatacctct tatcttcctc ccacagctcc tgggcaacgt gctggtctgt gtgctggccc
     2041 atcactttgg caaagaattc accccaccag tgcaggctgc ctatcagaaa gtggtggctg
     2101 gtgtggctaa tgccctggcc cacaagtatc acatggcttc caaggtgtac gaccccgagc
     2161 aacgcaaacg catgatcact gggcctcagt ggtgggctcg ctgcaagcaa atgaacgtgc
     2221 tggactcctt catcaactac tatgattccg agaagcacgc cgagaacgcc gtgatttttc
     2281 tgcatggtaa cgctgcctcc agctacctgt ggaggcacgt cgtgcctcac atcgagcccg
     2341 tggctagatg catcatccct gatctgatcg gaatgggtaa gtccggcaag agcgggaatg
     2401 gctcatatcg cctcctggat cactacaagt acctcaccgc ttggttcgag ctgctgaacc
     2461 ttccaaagaa aatcatcttt gtgggccacg actggggggc ttgtctggcc tttcactact
     2521 cctacgagca ccaagacaag atcaaggcca tcgtccatgc tgagagtgtc gtggacgtga
     2581 tcgagtcctg ggacgagtgg cctgacatcg aggaggatat cgccctgatc aagagcgaag
     2641 agggcgagaa aatggtgctt gagaataact tcttcgtcga gaccatgctc ccaagcaaga
     2701 tcatgcggaa actggagcct gaggagttcg ctgcctacct ggagccattc aaggagaagg
     2761 gcgaggttag acggcctacc ctctcctggc ctcgcgagat ccctctcgtt aagggaggca
     2821 agcccgacgt cgtccagatt gtccgcaact acaacgccta ccttcgggcc agcgacgatc
     2881 tgcctaagat gttcatcgag tccgaccctg ggttcttttc caacgctatt gtcgagggag
     2941 ctaagaagtt ccctaacacc gagttcgtga aggtgaaggg cctccacttc agccaggagg
     3001 acgctccaga tgaaatgggt aagtacatca agagcttcgt ggagcgcgtg ctgaagaacg
     3061 agcagggtaa gcctatccct aaccctctcc tcggtctcga ttctacgtaa gcggccgctc
     3121 cgctttcttg ctgtccaatt tctattaaag gttcctttgt tccctaagtc caactaccaa
     3181 actgggtcta gctctagctg tagaaaacat gaggatcacc catgtctgct ggacgactgt
     3241 agaaaacatg aggatcaccc atgtctgctg tctagctgta gaaaacatga ggatcaccca
     3301 tgtctgctgg acgactgtag aaaacatgag gatcacccat gtctgctgtc tagctgtaga
     3361 aaacatgagg atcacccatg tctgctggac gactgtagaa aacatgagga tcacccatgt
     3421 ctgctgtcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc
     3481 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3541 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3601 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3661 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3721 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3781 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3841 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3901 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3961 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4021 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4081 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4141 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4381 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4441 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4501 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4561 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4621 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4681 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4741 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4801 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4861 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4921 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4981 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5041 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5101 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5161 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5221 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5281 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5341 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5401 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5461 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5521 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5581 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5641 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5701 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5761 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5821 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5881 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5941 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6001 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6061 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt cccctggaag
     6241 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6301 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6361 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6421 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6481 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6541 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6601 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6661 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6721 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6781 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6841 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6901 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6961 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7021 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7081 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7141 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7201 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7261 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7321 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7381 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7441 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7501 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7561 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7621 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7681 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7741 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7801 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7861 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7921 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7981 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     8041 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     8101 gtacgggcca gatatacgcg tt
//
