LOCUS       sequence-471828-        7930 bp    DNA     circular SYN 08-MAY-2025
DEFINITION  Express 3A4 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7930)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7930)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 8, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/3A4/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..7930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2055..2064
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2061..3710
                     /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             3714..3833
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    3885..4006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3896..3915)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3950..3969
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4200..4557
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4200..4391
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4223..4243)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4408..4543
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4470..4489
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4588..5625
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5604..5612
                     /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    5649..5697
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5853..5870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6024..6612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6104..6123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6812..7603,7604..7672))
                     /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    7777..7825
                     /note="synthetic polyadenylation signal"
     misc_feature    7839..7930
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7879..7898
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccg tagtcgttag aatctgaact tcctgaagtt catgtgcaaa
       61 gttgagttaa ttgtttaata ttcaacaagg attatgccag taagatggta ggaaaatatt
      121 agatatgtgt catcactgct ggtattattt aaactgcaac atattttagc tggctgctga
      181 tctcagccac catgcctgca ttttatctct gtctcgtggt ctgcaacctt ggaagctttg
      241 aacttagctc atagaatcct gggcatcaag aacatgtggt tctaatggct agatagggaa
      301 tgagagtaaa aggattttgc ccacggtcac gtgagtaaac aacagatttg gaggggtctg
      361 gactactgtg atgacttcat tctgacaata tgttccagtt gtcctttcat ttcctcctaa
      421 tcacatgtct ggtctgatct ggctgtttcc caccttccaa ttcctgcctt ctccaatgct
      481 cccttccgta ggtcactctg tggctcagag accctgctta gcaagcgccc aacctttcaa
      541 ttatttgttc agtaaaactt gaactcatgt ctccccttct tgataaaaag aaaatacgtt
      601 atgtaatgtc gggttactct ataactcttg tcctgtctct cggcaactac tgaactaact
      661 gttttcatat tgagcaaacg tttatggaag gactgccaag agtcaggtac taggcttggt
      721 aatattcccc gttctctcta gtcaaagcca acaccagcca gacttgcaga tctaggtccc
      781 aagcccactg cagatcacag gccagggtct ggtctcctct gagctccttt gggagggaaa
      841 gacagaatta ttaacaccca ttttgtagat taggcaactg aggctgagga agtttaaata
      901 actcagacag ggcctgcacg tcagtcatat tccaaggatc cctactcact gtcttctctc
      961 tacagaacga gatgtctctg gaggctagcg agatggttca ttcctttcat ttgattatca
     1021 aagaaactca tgtcccaatt aaaggtcata aagcccagtt tgtaaactga gatgatctca
     1081 gctgaatgaa cttgctgacc ctctgctttc ctccagcctc tcggtgccct tgaaatcatg
     1141 tcggttcaag cagcctcatg aggcattaca aagtttaatt atttcagtga ttattaaacc
     1201 ttgtcctgtg ttgaccccag gtgaatcaca agctgaactt ctgacaagaa caagctatca
     1261 tattcttttc aattacagaa aaaagtaagt taattgatag gatttttttt gtttaaaaaa
     1321 aatgttacta gttttgaaaa ggtaatatgt gcacatggta aacactaaga aggtataaga
     1381 gcataatgct tttatactac taagaataat gttttctcta agtttttttt ggtagatgct
     1441 ttcatcagat taagaaaatt ccctgctatt agttgttgaa ggtttttata tcataaatga
     1501 aagttgaata ttattatcat atattattaa tatattgtta ttgaactatc aaagcctttt
     1561 cctaaaacca ttgagatgat cttataacca ttctccttta acctgttgac gagatctgta
     1621 ggtgtggctt gttgggatga atttcaagta ttttggaatg aggacagcca tagagacaag
     1681 ggcaggagag aggcgattta atagatttta tgccaatggc tccacttgag tttctgataa
     1741 gaacccagaa cccttggact ccccagtaac attgattgag ttgtttatga tacctcatag
     1801 aatatgaact caaaggaggt cagtgagtgg tgtgtgtgtg attctttgcc aacttccaag
     1861 gtggagaagc ctcttccaac tgcaggcaga gcacaggtgg ccctgctact ggctgcagct
     1921 ccagccctgc ctccttctct agcatataaa caatccaaca gcctcactga atcactgctg
     1981 tgcagggcag gaaagctcca tgcacatagc ccagcaaaga gcaacaagct tggcaatccg
     2041 gtactgttgg taaagccacc atggaagatg ccaaaaacat taagaagggc ccagcgccat
     2101 tctacccact cgaagacggg accgccggcg agcagctgca caaagccatg aagcgctacg
     2161 ccctggtgcc cggcaccatc gcctttaccg acgcacatat cgaggtggac attacctacg
     2221 ccgagtactt cgagatgagc gttcggctgg cagaagctat gaagcgctat gggctgaata
     2281 caaaccatcg gatcgtggtg tgcagcgaga atagcttgca gttcttcatg cccgtgttgg
     2341 gtgccctgtt catcggtgtg gctgtggccc cagctaacga catctacaac gagcgcgagc
     2401 tgctgaacag catgggcatc agccagccca ccgtcgtatt cgtgagcaag aaagggctgc
     2461 aaaagatcct caacgtgcaa aagaagctac cgatcataca aaagatcatc atcatggata
     2521 gcaagaccga ctaccagggc ttccaaagca tgtacacctt cgtgacttcc catttgccac
     2581 ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa accatcgccc
     2641 tgatcatgaa cagtagtggc agtaccggat tgcccaaggg cgtagcccta ccgcaccgca
     2701 ccgcttgtgt ccgattcagt catgcccgcg accccatctt cggcaaccag atcatccccg
     2761 acaccgctat cctcagcgtg gtgccatttc accacggctt cggcatgttc accacgctgg
     2821 gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag gagctattct
     2881 tgcgcagctt gcaagactat aagattcaat ctgccctgct ggtgcccaca ctatttagct
     2941 tcttcgctaa gagcactctc atcgacaagt acgacctaag caacttgcac gagatcgcca
     3001 gcggcggggc gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc ttccacctac
     3061 caggcatccg ccagggctac ggcctgacag aaacaaccag cgccattctg atcacccccg
     3121 aaggggacga caagcctggc gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg
     3181 tggacttgga caccggtaag acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg
     3241 gccccatgat catgagcggc tacgttaaca accccgaggc tacaaacgct ctcatcgaca
     3301 aggacggctg gctgcacagc ggcgacatcg cctactggga cgaggacgag cacttcttca
     3361 tcgtggaccg gctgaagagc ctgatcaaat acaagggcta ccaggtagcc ccagccgaac
     3421 tggagagcat cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg
     3481 acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga
     3541 ccgagaagga gatcgtggac tatgtggcca gccaggttac aaccgccaag aagctgcgcg
     3601 gtggtgttgt gttcgtggac gaggtgccta aaggactgac cggcaagttg gacgcccgca
     3661 agatccgcga gattctcatt aaggccaaga agggcggcaa gatcgccgtg aattctcacg
     3721 gcttccctcc cgaggtggag gagcaggccg ccggcaccct gcccatgagc tgcgcccagg
     3781 agagcggcat ggatagacac cctgctgctt gcgccagcgc caggatcaac gtctaaggcc
     3841 gcgactctag agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga
     3901 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3961 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4021 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     4081 cctctacaaa tgtggtaaaa tcgataagga tccgtttgcg tattgggcgc tcttccgctg
     4141 atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt agctaccttc
     4201 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4261 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4321 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4381 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4441 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4501 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4561 ctcgattctt ctgacactag cgccaccatg aagaagcccg aactcaccgc taccagcgtt
     4621 gaaaaatttc tcatcgagaa gttcgacagt gtgagcgacc tgatgcagtt gtcggagggc
     4681 gaagagagcc gagccttcag cttcgatgtc ggcggacgcg gctatgtact gcgggtgaat
     4741 agctgcgctg atggcttcta caaagaccgc tacgtgtacc gccacttcgc cagcgctgca
     4801 ctacccatcc ccgaagtgtt ggacatcggc gagttcagcg agagcctgac atactgcatc
     4861 agtagacgcg cccaaggcgt tactctccaa gacctccccg aaacagagct gcctgctgtg
     4921 ttacagcctg tcgccgaagc tatggatgct attgccgccg ccgacctcag tcaaaccagc
     4981 ggcttcggcc cattcgggcc ccaaggcatc ggccagtaca caacctggcg ggatttcatt
     5041 tgcgccattg ctgatcccca tgtctaccac tggcagaccg tgatggacga caccgtgtcc
     5101 gccagcgtag ctcaagccct ggacgaactg atgctgtggg ccgaagactg tcccgaggtg
     5161 cgccacctcg tccatgccga cttcggcagc aacaacgtcc tgaccgacaa cggccgcatc
     5221 accgccgtaa tcgactggtc cgaagctatg ttcggggaca gtcagtacga ggtggccaac
     5281 atcttcttct ggcggccctg gctggcttgc atggagcagc agactcgcta cttcgagcgc
     5341 cggcatcccg agctggccgg cagccctcgt ctgcgagcct acatgctgcg catcggcctg
     5401 gatcagctct accagagcct cgtggacggc aacttcgacg atgctgcctg ggctcaaggc
     5461 cgctgcgatg ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca aatcgctcgc
     5521 cggagcgcag ccgtatggac cgacggctgc gtcgaggtgc tggccgacag cggcaaccgc
     5581 cggcccagta cacgaccgcg cgctaaggag gtaggtcgag tttaaactct agaaccggtc
     5641 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgttc
     5701 gaactagatg ctgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     5761 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     5821 taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5881 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5941 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6001 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6421 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6781 agcggccgca aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga
     6841 tctcagcgat ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca
     6901 ctacgattcg tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc
     6961 gttcaccggc ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa
     7021 gtggtcctgc tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag
     7081 taagaagttc gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg
     7141 tatcacgctc gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg
     7201 tcacatgatc acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg
     7261 tcagaagtaa gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc
     7321 ttaccgtcat gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt
     7381 tttgtgagta gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca
     7441 ccgcgccaca tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga
     7501 aagactcaag gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca
     7561 gttgatcttc agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc
     7621 aaaatgccgc aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc
     7681 tttttcaata ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa
     7741 gtaatattaa ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta
     7801 catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac
     7861 aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga
     7921 acatttctct
//
