LOCUS       sequence-83025-e        6208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dicistronic vector with EMCV IRES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6208)
  AUTHORS   Burman LG, Mauro VP
  TITLE     Analysis of rRNA processing and translation in mammalian cells using
            a synthetic 18S rRNA expression system.
  JOURNAL   Nucleic Acids Res. 2012 Sep;40(16):8085-98. doi: 10.1093/nar/gks530.
            Epub 2012 Jun 20.
  PUBMED    22718970
REFERENCE   2  (bases 1 to 6208)
  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"; doi: 
            "10.1093/nar/gks530"; journalName: "Nucleic Acids Res"; date: 
            "2012-09"; volume: "40"; issue: "16"; pages: "8085-98"
COMMENT     Sequence Label: R5 EMCV
FEATURES             Location/Qualifiers
     source          1..6208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        29..386
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        29..220
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(52..72)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      237..372
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      416..425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             422..2074
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2091..2107)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2092..2108)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    2128..2695
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2289..2306)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2516..2535
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2709..3644
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3685..3806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3696..3715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3750..3769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4054..4071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4225..4813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4305..4324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5013..5804,5805..5873))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5978..6026
                     /note="synthetic polyadenylation signal"
     misc_feature    6040..6131
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6080..6099
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
       61 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      121 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
      181 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
      241 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
      301 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
      361 tttttggagg cctaggcttt tgcaaaaagc ttggcaatcc ggtactgttg gtaaagccac
      421 catggaagat gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg
      481 gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat
      541 cgcctttacc gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag
      601 cgttcggctg gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt
      661 gtgcagcgag aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt
      721 ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat
      781 cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca
      841 aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg
      901 cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga
      961 cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg
     1021 cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag
     1081 tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt
     1141 ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt
     1201 tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta
     1261 taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct
     1321 catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag
     1381 caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta
     1441 cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg
     1501 cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa
     1561 gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg
     1621 ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag
     1681 cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag
     1741 cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca
     1801 acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct
     1861 gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga
     1921 ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga
     1981 cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat
     2041 taaggccaag aagggcggca agatcgccgt gtaataattc tagagcttat cgataccgtc
     2101 gacctcgaat cactagtcag ctggaattcc gccccccccc cctaacgtta ctggccgaag
     2161 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     2221 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     2281 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     2341 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     2401 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     2461 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     2521 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     2581 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     2641 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg
     2701 ccacaaccat ggcttccaag gtgtacgacc ccgagcaacg caaacgcatg atcactgggc
     2761 ctcagtggtg ggctcgctgc aagcaaatga acgtgctgga ctccttcatc aactactatg
     2821 attccgagaa gcacgccgag aacgccgtga tttttctgca tggtaacgct gcctccagct
     2881 acctgtggag gcacgtcgtg cctcacatcg agcccgtggc tagatgcatc atccctgatc
     2941 tgatcggaat gggtaagtcc ggcaagagcg ggaatggctc atatcgcctc ctggatcact
     3001 acaagtacct caccgcttgg ttcgagctgc tgaaccttcc aaagaaaatc atctttgtgg
     3061 gccacgactg gggggcttgt ctggcctttc actactccta cgagcaccaa gacaagatca
     3121 aggccatcgt ccatgctgag agtgtcgtgg acgtgatcga gtcctgggac gagtggcctg
     3181 acatcgagga ggatatcgcc ctgatcaaga gcgaagaggg cgagaaaatg gtgcttgaga
     3241 ataacttctt cgtcgagacc atgctcccaa gcaagatcat gcggaaactg gagcctgagg
     3301 agttcgctgc ctacctggag ccattcaagg agaagggcga ggttagacgg cctaccctct
     3361 cctggcctcg cgagatccct ctcgttaagg gaggcaagcc cgacgtcgtc cagattgtcc
     3421 gcaactacaa cgcctacctt cgggccagcg acgatctgcc taagatgttc atcgagtccg
     3481 accctgggtt cttttccaac gctattgtcg agggagctaa gaagttccct aacaccgagt
     3541 tcgtgaaggt gaagggcctc cacttcagcc aggaggacgc tccagatgaa atgggtaagt
     3601 acatcaagag cttcgtggag cgcgtgctga agaacgagca gtaattctag agtcggggcg
     3661 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3721 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3781 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3841 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3901 tcgataagga tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     3961 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     4021 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4861 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4921 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4981 cagcggccgc aaatgctaaa ccactgcagt ggttaccagt gcttgatcag tgaggcaccg
     5041 atctcagcga tctgcctatt tcgttcgtcc atagtggcct gactccccgt cgtgtagatc
     5101 actacgattc gtgagggctt accatcaggc cccagcgcag caatgatgcc gcgagagccg
     5161 cgttcaccgg cccccgattt gtcagcaatg aaccagccag cagggagggc cgagcgaaga
     5221 agtggtcctg ctactttgtc cgcctccatc cagtctatga gctgctgtcg tgatgctaga
     5281 gtaagaagtt cgccagtgag tagtttccga agagttgtgg ccattgctac tggcatcgtg
     5341 gtatcacgct cgtcgttcgg tatggcttcg ttcaactctg gttcccagcg gtcaagccgg
     5401 gtcacatgat cacccatatt atgaagaaat gcagtcagct ccttagggcc tccgatcgtt
     5461 gtcagaagta agttggccgc ggtgttgtcg ctcatggtaa tggcagcact acacaattct
     5521 cttaccgtca tgccatccgt aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg
     5581 ttttgtgagt agtgtatacg gcgaccaagc tgctcttgcc cggcgtctat acgggacaac
     5641 accgcgccac atagcagtac tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg
     5701 aaagactcaa ggatcttgcc gctattgaga tccagttcga tatagcccac tcttgcaccc
     5761 agttgatctt cagcatcttt tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag
     5821 caaaatgccg caaagaaggg aatgagtgcg acacgaaaat gttggatgct catactcgtc
     5881 ctttttcaat attattgaag catttatcag ggttactagt acgtctctca aggataagta
     5941 agtaatatta aggtacggga ggtattggac aggccgcaat aaaatatctt tattttcatt
     6001 acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa
     6061 caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag
     6121 aacatttctc tggcctaact ggccggtacc tgagctcgct agcctcgagg atatcagatc
     6181 tgcgcagcac catggcctga aataacct
//
