LOCUS       sequence-83027-e        6396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dicistronic vector with PV IRES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6396)
  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 6396)
  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 PV
FEATURES             Location/Qualifiers
     source          1..6396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        56..247
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(79..99)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      264..399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      443..452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             449..2101
                     /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(2118..2134)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2119..2135)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        2163..2181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2924..3859
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3900..4021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3911..3930)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3965..3984
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4269..4286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4440..5028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4520..4539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5228..6019,6020..6088))
                     /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    6193..6241
                     /note="synthetic polyadenylation signal"
     misc_feature    6255..6346
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6295..6314
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
       61 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
      301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
      361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
      421 gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta agaagggccc
      481 agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca aagccatgaa
      541 gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg aggtggacat
      601 tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga agcgctatgg
      661 gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt tcttcatgcc
      721 cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca tctacaacga
      781 gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg tgagcaagaa
      841 agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa agatcatcat
      901 catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg tgacttccca
      961 tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc gggacaaaac
     1021 catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg tagccctacc
     1081 gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg gcaaccagat
     1141 catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg gcatgttcac
     1201 cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct tcgaggagga
     1261 gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg tgcccacact
     1321 atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca acttgcacga
     1381 gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg ccaaacgctt
     1441 ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg ccattctgat
     1501 cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct tcttcgaggc
     1561 taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg gcgagctgtg
     1621 cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta caaacgctct
     1681 catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg aggacgagca
     1741 cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc aggtagcccc
     1801 agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg gggtcgccgg
     1861 cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg aacacggtaa
     1921 aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa ccgccaagaa
     1981 gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg gcaagttgga
     2041 cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga tcgccgtgta
     2101 ataattctag agcttatcga taccgtcgac ctcgaatcac tagtcagctg gcttatcgaa
     2161 attaatacga ctcactatag gttaaaacag ctctggggtt gtacccaccc cagaggccca
     2221 cgtggcggct agtactccgg tattgcggta cccttgtacg cctgttttat actcccttcc
     2281 cgtaacttag acgcacaaaa ccaagttcaa tagaaggggg tacaaaccag taccaccacg
     2341 aacaagcact tctgtttccc cggtgatgtc gtatagactg cttgcgtggt tgaaagcgac
     2401 ggatccgtta tccgcttatg tacttcgaga agcccagtac cacctcggaa tcttcgatgc
     2461 gttgcgctca gcactcaacc ccagagtgta gcttaggctg atgagtctgg acatccctca
     2521 ccggtgacgg tggtccaggc tgcgttggcg gcctacctat ggctaacgcc atgggacgct
     2581 agttgtgaac aaggtgtgaa gagcctattg agctacataa gaatcctccg gcccctgaat
     2641 gcggctaatc ccaacctcgg agcaggtggt cacaaaccag tgattggcct gtcgtaacgc
     2701 gcaagtccgt ggcggaaccg actactttgg gtgtccgtgt ttccttttat tttattgtgg
     2761 ctgcttatgg tgacaatcac agattgttat cataaagcga attggattgg ccatccggtg
     2821 aaagtgagac tcattatcta tctgtttgct ggatccgctc cattgagtgt gtttactcta
     2881 agtacaattt caacagttat ttcaatcaga caattgtatc accatggctt ccaaggtgta
     2941 cgaccccgag caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca
     3001 aatgaacgtg ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc
     3061 cgtgattttt ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca
     3121 catcgagccc gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa
     3181 gagcgggaat ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga
     3241 gctgctgaac cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc
     3301 ctttcactac tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt
     3361 cgtggacgtg atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat
     3421 caagagcgaa gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct
     3481 cccaagcaag atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt
     3541 caaggagaag ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt
     3601 taagggaggc aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc
     3661 cagcgacgat ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat
     3721 tgtcgaggga gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt
     3781 cagccaggag gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt
     3841 gctgaagaac gagcagtaat tctagagtcg gggcggccgg ccgcttcgag cagacatgat
     3901 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3961 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4021 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4081 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt cgaccgatgc
     4141 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     4201 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     4261 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4321 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4381 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4441 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4501 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4561 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4621 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4681 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4741 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4801 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4861 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4921 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4981 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5041 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5101 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5161 tcaatctaaa gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact
     5221 gcagtggtta ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt
     5281 cgtccatagt ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat
     5341 caggccccag cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag
     5401 caatgaacca gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct
     5461 ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt
     5521 tccgaagagt tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg
     5581 cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa
     5641 gaaatgcagt cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt
     5701 tgtcgctcat ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat
     5761 gcttttccgt gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac
     5821 caagctgctc ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga
     5881 aagtgctcat catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat
     5941 tgagatccag ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt
     6001 tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga
     6061 gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt
     6121 atcagggtta ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat
     6181 tggacaggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     6241 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     6301 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctggcc taactggccg
     6361 gtacctgagc tcgctagcct cgaggatatc agatct
//
