LOCUS       sequence-472435-        6267 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express RBP-Jk response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/RBP-Jk-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        335..366
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      393..402
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             399..2048
                     /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"
     CDS             2052..2171
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2223..2344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2234..2253)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2288..2307
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2538..2895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2538..2729
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2561..2581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2746..2881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2808..2827
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2926..3963
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3942..3950
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3987..4035
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4191..4208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4362..4950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4442..4461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5150..5941,5942..6010))
                     /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    6115..6163
                     /note="synthetic polyadenylation signal"
     primer_bind     6217..6236
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctccgac tcgtgggaaa atgggcggaa gggcaccgtg
       61 ggaaaatagt cgactcgtgg gaaaatgggc ggaagggcac cgtgggaaaa tagtcgactc
      121 gtgggaaaat gggcggaagg gcaccgtggg aaaatagtcg actcgtggga aaatgggcgg
      181 aagggcaccg tgggaaaata gtcgactcgt gggaaaatgg gcggaagggc accgtgggaa
      241 aatagtcgac tcgtgggaaa atgggcggaa gggcaccgtg ggaaaatagt ctcgaggata
      301 tcaagatctg gcctcggcgg ccaagcttag acactagagg gtatataatg gaagctcgac
      361 ttccagcttg gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta
      421 agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca
      481 aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg
      541 aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga
      601 agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt
      661 tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca
      721 tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg
      781 tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa
      841 agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg
      901 tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc
      961 gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg
     1021 tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg
     1081 gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg
     1141 gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct
     1201 tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg
     1261 tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca
     1321 acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg
     1381 ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg
     1441 ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct
     1501 tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg
     1561 gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta
     1621 caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg
     1681 aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc
     1741 aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg
     1801 gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg
     1861 aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa
     1921 ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg
     1981 gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga
     2041 tcgccgtgaa ttctcacggc ttccctcccg aggtggagga gcaggccgcc ggcaccctgc
     2101 ccatgagctg cgcccaggag agcggcatgg atagacaccc tgctgcttgc gccagcgcca
     2161 ggatcaacgt ctaaggccgc gactctagag tcggggcggc cggccgcttc gagcagacat
     2221 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2281 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2341 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2401 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta
     2461 ttgggcgctc ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     2521 acttggttag ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     2581 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2641 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2701 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2761 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2821 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2881 ctaggctttt gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa
     2941 ctcaccgcta ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg
     3001 atgcagttgt cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc
     3061 tatgtactgc gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc
     3121 cacttcgcca gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag
     3181 agcctgacat actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa
     3241 acagagctgc ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc
     3301 gacctcagtc aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca
     3361 acctggcggg atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg
     3421 atggacgaca ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc
     3481 gaagactgtc ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg
     3541 accgacaacg gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt
     3601 cagtacgagg tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag
     3661 actcgctact tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac
     3721 atgctgcgca tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat
     3781 gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt
     3841 cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg
     3901 gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt
     3961 taaactctag aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg
     4021 ttggtttttt gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc
     4081 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     4141 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     4201 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4261 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4321 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4801 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4921 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4981 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5041 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5101 gagtaaactt ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct
     5161 tgatcagtga ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac
     5221 tccccgtcgt gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa
     5281 tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag
     5341 ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct
     5401 gctgtcgtga tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca
     5461 ttgctactgg catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt
     5521 cccagcggtc aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct
     5581 tagggcctcc gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg
     5641 cagcactaca caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg
     5701 agtactcaac caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg
     5761 cgtctatacg ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga
     5821 atcgttcttc ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat
     5881 agcccactct tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt
     5941 gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt
     6001 ggatgctcat actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg
     6061 tctctcaagg ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa
     6121 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
     6181 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
     6241 gcaagtgcag gtgccagaac atttctc
//
