LOCUS       sequence-357810-        6672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of K0-Rluc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6672)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6672)
  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"
COMMENT     Sequence Label: pEGFP-N3-FLuc-K0-Rluc_AH
FEATURES             Location/Qualifiers
     source          1..6672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             606..2255
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEYAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(672..690)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             2262..2303
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2313..2369
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2370..2396
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2403..3338
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3460..3581
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3497..3516)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3551..3570
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3588..4043)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3725..3746)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3937..3956
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4070..4174
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4176..4533
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4176..4367
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4199..4219)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4384..4519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4568..5362
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4568..5362
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4622..4641)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5232..5251
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5550..5569)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5594..5641
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5970..6558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6459..6478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacag
      601 ccaccatgga atacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag
      661 atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa
      721 caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa
      781 tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg
      841 tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg
      901 gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg
      961 gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg
     1021 tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc
     1081 agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat
     1141 acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct
     1201 ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat
     1261 tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa
     1321 gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg
     1381 gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg
     1441 attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca
     1501 ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc
     1561 tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag
     1621 gatatgggct cactgagact acatcagcta ttctgattac acccgagggg gatgataaac
     1681 cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg
     1741 ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt
     1801 ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac
     1861 attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga
     1921 agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc
     1981 tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg
     2041 aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg
     2101 tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg
     2161 tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc
     2221 tcataaaggc caagaagggc ggaaagatcg ccgtgggatc cggtaagcct atccctaacc
     2281 ctctcctcgg tctcgattct acgggaagcg gagctactaa cttcagcctg ctgaagcagg
     2341 ctggagacgt ggaggagaac cctggacctt acccatacga tgttccagat tacgctgaat
     2401 tcatggcttc caaggtgtac gaccccgagc aacgcaaacg catgatcact gggcctcagt
     2461 ggtgggctcg ctgcaagcaa atgaacgtgc tggactcctt catcaactac tatgattccg
     2521 agaagcacgc cgagaacgcc gtgatttttc tgcatggtaa cgctgcctcc agctacctgt
     2581 ggaggcacgt cgtgcctcac atcgagcccg tggctagatg catcatccct gatctgatcg
     2641 gaatgggtaa gtccggcaag agcgggaatg gctcatatcg cctcctggat cactacaagt
     2701 acctcaccgc ttggttcgag ctgctgaacc ttccaaagaa aatcatcttt gtgggccacg
     2761 actggggggc ttgtctggcc tttcactact cctacgagca ccaagacaag atcaaggcca
     2821 tcgtccatgc tgagagtgtc gtggacgtga tcgagtcctg ggacgagtgg cctgacatcg
     2881 aggaggatat cgccctgatc aagagcgaag agggcgagaa aatggtgctt gagaataact
     2941 tcttcgtcga gaccatgctc ccaagcaaga tcatgcggaa actggagcct gaggagttcg
     3001 ctgcctacct ggagccattc aaggagaagg gcgaggttag acggcctacc ctctcctggc
     3061 ctcgcgagat ccctctcgtt aagggaggca agcccgacgt cgtccagatt gtccgcaact
     3121 acaacgccta ccttcgggcc agcgacgatc tgcctaagat gttcatcgag tccgaccctg
     3181 ggttcttttc caacgctatt gtcgagggag ctaagaagtt ccctaacacc gagttcgtga
     3241 aggtgaaggg cctccacttc agccaggagg acgctccaga tgaaatgggt aagtacatca
     3301 agagcttcgt ggagcgcgtg ctgaagaacg agcagtaagc ggccgcgact ctagatcata
     3361 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3421 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3481 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3541 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     3601 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3661 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3721 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3781 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3841 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3901 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3961 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4021 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4081 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4141 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4201 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4261 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4321 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4381 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4441 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4501 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4561 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4621 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4681 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4741 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4801 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4861 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4921 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4981 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5041 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     5101 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5161 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5221 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5281 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5341 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5401 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5461 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5521 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5581 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5641 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5701 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5761 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5821 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5881 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6601 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6661 accgccatgc at
//
