LOCUS       sequence-83028-e        5644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dicistronic vector with cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5644)
  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 5644)
  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 MCS
FEATURES             Location/Qualifiers
     source          1..5644
                     /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"
     regulatory      2166..2175
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2172..3107
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3148..3269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3159..3178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3213..3232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3517..3534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3688..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3768..3787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4476..5267,5268..5336))
                     /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    5441..5489
                     /note="synthetic polyadenylation signal"
     misc_feature    5503..5594
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5543..5562
                     /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 gaattcgttg
     2161 gtaaagccac catggcttcc aaggtgtacg accccgagca acgcaaacgc atgatcactg
     2221 ggcctcagtg gtgggctcgc tgcaagcaaa tgaacgtgct ggactccttc atcaactact
     2281 atgattccga gaagcacgcc gagaacgccg tgatttttct gcatggtaac gctgcctcca
     2341 gctacctgtg gaggcacgtc gtgcctcaca tcgagcccgt ggctagatgc atcatccctg
     2401 atctgatcgg aatgggtaag tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc
     2461 actacaagta cctcaccgct tggttcgagc tgctgaacct tccaaagaaa atcatctttg
     2521 tgggccacga ctggggggct tgtctggcct ttcactactc ctacgagcac caagacaaga
     2581 tcaaggccat cgtccatgct gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc
     2641 ctgacatcga ggaggatatc gccctgatca agagcgaaga gggcgagaaa atggtgcttg
     2701 agaataactt cttcgtcgag accatgctcc caagcaagat catgcggaaa ctggagcctg
     2761 aggagttcgc tgcctacctg gagccattca aggagaaggg cgaggttaga cggcctaccc
     2821 tctcctggcc tcgcgagatc cctctcgtta agggaggcaa gcccgacgtc gtccagattg
     2881 tccgcaacta caacgcctac cttcgggcca gcgacgatct gcctaagatg ttcatcgagt
     2941 ccgaccctgg gttcttttcc aacgctattg tcgagggagc taagaagttc cctaacaccg
     3001 agttcgtgaa ggtgaagggc ctccacttca gccaggagga cgctccagat gaaatgggta
     3061 agtacatcaa gagcttcgtg gagcgcgtgc tgaagaacga gcagtaattc tagagtcggg
     3121 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3181 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3241 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3301 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3361 aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     3421 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     3481 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagcggc cgcaaatgct aaaccactgc agtggttacc agtgcttgat cagtgaggca
     4501 ccgatctcag cgatctgcct atttcgttcg tccatagtgg cctgactccc cgtcgtgtag
     4561 atcactacga ttcgtgaggg cttaccatca ggccccagcg cagcaatgat gccgcgagag
     4621 ccgcgttcac cggcccccga tttgtcagca atgaaccagc cagcagggag ggccgagcga
     4681 agaagtggtc ctgctacttt gtccgcctcc atccagtcta tgagctgctg tcgtgatgct
     4741 agagtaagaa gttcgccagt gagtagtttc cgaagagttg tggccattgc tactggcatc
     4801 gtggtatcac gctcgtcgtt cggtatggct tcgttcaact ctggttccca gcggtcaagc
     4861 cgggtcacat gatcacccat attatgaaga aatgcagtca gctccttagg gcctccgatc
     4921 gttgtcagaa gtaagttggc cgcggtgttg tcgctcatgg taatggcagc actacacaat
     4981 tctcttaccg tcatgccatc cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag
     5041 tcgttttgtg agtagtgtat acggcgacca agctgctctt gcccggcgtc tatacgggac
     5101 aacaccgcgc cacatagcag tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg
     5161 cggaaagact caaggatctt gccgctattg agatccagtt cgatatagcc cactcttgca
     5221 cccagttgat cttcagcatc ttttactttc accagcgttt cggggtgtgc aaaaacaggc
     5281 aagcaaaatg ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat gctcatactc
     5341 gtcctttttc aatattattg aagcatttat cagggttact agtacgtctc tcaaggataa
     5401 gtaagtaata ttaaggtacg ggaggtattg gacaggccgc aataaaatat ctttattttc
     5461 attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca
     5521 aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc
     5581 cagaacattt ctctggccta actggccggt acctgagctc gctagcctcg aggatatcag
     5641 atct
//
