LOCUS       sequence-471820-        6060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GAS response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6060)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6060)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pGL4 [luc2P/GAS-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..158
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      185..194
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             191..1840
                     /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             1844..1963
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2015..2136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2026..2045)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2080..2099
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2330..2687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2330..2521
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2353..2373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2538..2673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2600..2619
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2718..3755
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3734..3742
                     /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    3779..3827
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3983..4000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4154..4742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4234..4253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4942..5733,5734..5802))
                     /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    5907..5955
                     /note="synthetic polyadenylation signal"
     misc_feature    5969..6060
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6009..6028
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcagtt tcatattact ctaaatcagt ttcatattac
       61 tctaaatcag tttcatatta ctctaaatca gtttcatatt actctaaatc ctcgaagctt
      121 agacactaga gggtatataa tggaagctcg acttccagct tggcaatccg gtactgttgg
      181 taaagccacc atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact
      241 cgaagacggg accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc
      301 cggcaccatc gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt
      361 cgagatgagc gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg
      421 gatcgtggtg tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt
      481 catcggtgtg gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag
      541 catgggcatc agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct
      601 caacgtgcaa aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga
      661 ctaccagggc ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa
      721 cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa
      781 cagtagtggc agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt
      841 ccgattcagt catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat
      901 cctcagcgtg gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat
      961 ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt
     1021 gcaagactat aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa
     1081 gagcactctc atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc
     1141 gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg
     1201 ccagggctac ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga
     1261 caagcctggc gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga
     1321 caccggtaag acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat
     1381 catgagcggc tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg
     1441 gctgcacagc ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg
     1501 gctgaagagc ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat
     1561 cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc
     1621 cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga
     1681 gatcgtggac tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt
     1741 gttcgtggac gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga
     1801 gattctcatt aaggccaaga agggcggcaa gatcgccgtg aattctcacg gcttccctcc
     1861 cgaggtggag gagcaggccg ccggcaccct gcccatgagc tgcgcccagg agagcggcat
     1921 ggatagacac cctgctgctt gcgccagcgc caggatcaac gtctaaggcc gcgactctag
     1981 agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2041 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2101 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2161 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2221 tgtggtaaaa tcgataagga tccgtttgcg tattgggcgc tcttccgctg atctgcgcag
     2281 caccatggcc tgaaataacc tctgaaagag gaacttggtt agctaccttc tgaggcggaa
     2341 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2401 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2521 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2581 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2641 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcgattctt
     2701 ctgacactag cgccaccatg aagaagcccg aactcaccgc taccagcgtt gaaaaatttc
     2761 tcatcgagaa gttcgacagt gtgagcgacc tgatgcagtt gtcggagggc gaagagagcc
     2821 gagccttcag cttcgatgtc ggcggacgcg gctatgtact gcgggtgaat agctgcgctg
     2881 atggcttcta caaagaccgc tacgtgtacc gccacttcgc cagcgctgca ctacccatcc
     2941 ccgaagtgtt ggacatcggc gagttcagcg agagcctgac atactgcatc agtagacgcg
     3001 cccaaggcgt tactctccaa gacctccccg aaacagagct gcctgctgtg ttacagcctg
     3061 tcgccgaagc tatggatgct attgccgccg ccgacctcag tcaaaccagc ggcttcggcc
     3121 cattcgggcc ccaaggcatc ggccagtaca caacctggcg ggatttcatt tgcgccattg
     3181 ctgatcccca tgtctaccac tggcagaccg tgatggacga caccgtgtcc gccagcgtag
     3241 ctcaagccct ggacgaactg atgctgtggg ccgaagactg tcccgaggtg cgccacctcg
     3301 tccatgccga cttcggcagc aacaacgtcc tgaccgacaa cggccgcatc accgccgtaa
     3361 tcgactggtc cgaagctatg ttcggggaca gtcagtacga ggtggccaac atcttcttct
     3421 ggcggccctg gctggcttgc atggagcagc agactcgcta cttcgagcgc cggcatcccg
     3481 agctggccgg cagccctcgt ctgcgagcct acatgctgcg catcggcctg gatcagctct
     3541 accagagcct cgtggacggc aacttcgacg atgctgcctg ggctcaaggc cgctgcgatg
     3601 ccatcgtccg cagcggggcc ggcaccgtcg gtcgcacaca aatcgctcgc cggagcgcag
     3661 ccgtatggac cgacggctgc gtcgaggtgc tggccgacag cggcaaccgc cggcccagta
     3721 cacgaccgcg cgctaaggag gtaggtcgag tttaaactct agaaccggtc atggccgcaa
     3781 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgttc gaactagatg
     3841 ctgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3901 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3961 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4021 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agcggccgca
     4921 aatgctaaac cactgcagtg gttaccagtg cttgatcagt gaggcaccga tctcagcgat
     4981 ctgcctattt cgttcgtcca tagtggcctg actccccgtc gtgtagatca ctacgattcg
     5041 tgagggctta ccatcaggcc ccagcgcagc aatgatgccg cgagagccgc gttcaccggc
     5101 ccccgatttg tcagcaatga accagccagc agggagggcc gagcgaagaa gtggtcctgc
     5161 tactttgtcc gcctccatcc agtctatgag ctgctgtcgt gatgctagag taagaagttc
     5221 gccagtgagt agtttccgaa gagttgtggc cattgctact ggcatcgtgg tatcacgctc
     5281 gtcgttcggt atggcttcgt tcaactctgg ttcccagcgg tcaagccggg tcacatgatc
     5341 acccatatta tgaagaaatg cagtcagctc cttagggcct ccgatcgttg tcagaagtaa
     5401 gttggccgcg gtgttgtcgc tcatggtaat ggcagcacta cacaattctc ttaccgtcat
     5461 gccatccgta agatgctttt ccgtgaccgg cgagtactca accaagtcgt tttgtgagta
     5521 gtgtatacgg cgaccaagct gctcttgccc ggcgtctata cgggacaaca ccgcgccaca
     5581 tagcagtact ttgaaagtgc tcatcatcgg gaatcgttct tcggggcgga aagactcaag
     5641 gatcttgccg ctattgagat ccagttcgat atagcccact cttgcaccca gttgatcttc
     5701 agcatctttt actttcacca gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc
     5761 aaagaaggga atgagtgcga cacgaaaatg ttggatgctc atactcgtcc tttttcaata
     5821 ttattgaagc atttatcagg gttactagta cgtctctcaa ggataagtaa gtaatattaa
     5881 ggtacgggag gtattggaca ggccgcaata aaatatcttt attttcatta catctgtgtg
     5941 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
     6001 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
//
