LOCUS       sequence-492597-        6946 bp    DNA     circular SYN 11-NOV-2025
DEFINITION  Express hEGR1 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6946)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 11, 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            "Unpublished"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/hEGR1/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    916..964
                     /note="synthetic polyadenylation signal"
     CDS             complement(988..2025)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             1001..1009
                     /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)"
     promoter        complement(2056..2413)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2070..2205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2124..2143)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        2222..2413
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2370..2390
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2607..2728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2644..2663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2698..2717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2780..2899)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(2903..4552)
                     /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"
     regulatory      4549..4558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    5629..5720
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(5661..5680)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    5734..5782
                     /note="synthetic polyadenylation signal"
     CDS             join(5887..5955,5956..6747)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgctgccg
      781 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
      841 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
      901 agcatctagt tcgaacacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
      961 tattgcggcc atgaccggtt ctagagttta aactcgacct acctccttag cgcgcggtcg
     1021 tgtactgggc cggcggttgc cgctgtcggc cagcacctcg acgcagccgt cggtccatac
     1081 ggctgcgctc cggcgagcga tttgtgtgcg accgacggtg ccggccccgc tgcggacgat
     1141 ggcatcgcag cggccttgag cccaggcagc atcgtcgaag ttgccgtcca cgaggctctg
     1201 gtagagctga tccaggccga tgcgcagcat gtaggctcgc agacgagggc tgccggccag
     1261 ctcgggatgc cggcgctcga agtagcgagt ctgctgctcc atgcaagcca gccagggccg
     1321 ccagaagaag atgttggcca cctcgtactg actgtccccg aacatagctt cggaccagtc
     1381 gattacggcg gtgatgcggc cgttgtcggt caggacgttg ttgctgccga agtcggcatg
     1441 gacgaggtgg cgcacctcgg gacagtcttc ggcccacagc atcagttcgt ccagggcttg
     1501 agctacgctg gcggacacgg tgtcgtccat cacggtctgc cagtggtaga catggggatc
     1561 agcaatggcg caaatgaaat cccgccaggt tgtgtactgg ccgatgcctt ggggcccgaa
     1621 tgggccgaag ccgctggttt gactgaggtc ggcggcggca atagcatcca tagcttcggc
     1681 gacaggctgt aacacagcag gcagctctgt ttcggggagg tcttggagag taacgccttg
     1741 ggcgcgtcta ctgatgcagt atgtcaggct ctcgctgaac tcgccgatgt ccaacacttc
     1801 ggggatgggt agtgcagcgc tggcgaagtg gcggtacacg tagcggtctt tgtagaagcc
     1861 atcagcgcag ctattcaccc gcagtacata gccgcgtccg ccgacatcga agctgaaggc
     1921 tcggctctct tcgccctccg acaactgcat caggtcgctc acactgtcga acttctcgat
     1981 gagaaatttt tcaacgctgg tagcggtgag ttcgggcttc ttcatggtgg cgctagtgtc
     2041 agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     2101 gaatagctca gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     2161 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     2221 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     2281 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     2341 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     2401 ctttccgcct cagaaggtag ctaaccaagt tcctctttca gaggttattt caggccatgg
     2461 tgctgcgcag atcagcggaa gagcgcccaa tacgcaaacg gatccttatc gattttacca
     2521 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2581 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2641 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2701 gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg gccgccccga
     2761 ctctagagtc gcggccttag acgttgatcc tggcgctggc gcaagcagca gggtgtctat
     2821 ccatgccgct ctcctgggcg cagctcatgg gcagggtgcc ggcggcctgc tcctccacct
     2881 cgggagggaa gccgtgagaa ttcacggcga tcttgccgcc cttcttggcc ttaatgagaa
     2941 tctcgcggat cttgcgggcg tccaacttgc cggtcagtcc tttaggcacc tcgtccacga
     3001 acacaacacc accgcgcagc ttcttggcgg ttgtaacctg gctggccaca tagtccacga
     3061 tctccttctc ggtcatggtt ttaccgtgtt ccagcacgac gactgcggcg ggcagctcgc
     3121 cggcatcgtc gtcgggcagg ccggcgaccc cggcgtcgaa gatgttgggg tgttgcagca
     3181 ggatgctctc cagttcggct ggggctacct ggtagccctt gtatttgatc aggctcttca
     3241 gccggtccac gatgaagaag tgctcgtcct cgtcccagta ggcgatgtcg ccgctgtgca
     3301 gccagccgtc cttgtcgatg agagcgtttg tagcctcggg gttgttaacg tagccgctca
     3361 tgatcatggg gccacggacg cacagctcgc cgcgctggtt cacacccagt gtcttaccgg
     3421 tgtccaagtc caccacctta gcctcgaaga agggcaccac cttgcctact gcgccaggct
     3481 tgtcgtcccc ttcgggggtg atcagaatgg cgctggttgt ttctgtcagg ccgtagccct
     3541 ggcggatgcc tggtaggtgg aagcgtttgg ccacggcctc acctacctcc ttgctgagcg
     3601 gcgccccgcc gctggcgatc tcgtgcaagt tgcttaggtc gtacttgtcg atgagagtgc
     3661 tcttagcgaa gaagctaaat agtgtgggca ccagcagggc agattgaatc ttatagtctt
     3721 gcaagctgcg caagaatagc tcctcctcga agcggtacat gagcacgacc cgaaagccgc
     3781 agatcaagta gcccagcgtg gtgaacatgc cgaagccgtg gtgaaatggc accacgctga
     3841 ggatagcggt gtcggggatg atctggttgc cgaagatggg gtcgcgggca tgactgaatc
     3901 ggacacaagc ggtgcggtgc ggtagggcta cgcccttggg caatccggta ctgccactac
     3961 tgttcatgat cagggcgatg gttttgtccc ggtcgaagct ctcgggcacg aagtcgtact
     4021 cgttgaagcc gggtggcaaa tgggaagtca cgaaggtgta catgctttgg aagccctggt
     4081 agtcggtctt gctatccatg atgatgatct tttgtatgat cggtagcttc ttttgcacgt
     4141 tgaggatctt ttgcagccct ttcttgctca cgaatacgac ggtgggctgg ctgatgccca
     4201 tgctgttcag cagctcgcgc tcgttgtaga tgtcgttagc tggggccaca gccacaccga
     4261 tgaacagggc acccaacacg ggcatgaaga actgcaagct attctcgctg cacaccacga
     4321 tccgatggtt tgtattcagc ccatagcgct tcatagcttc tgccagccga acgctcatct
     4381 cgaagtactc ggcgtaggta atgtccacct cgatatgtgc gtcggtaaag gcgatggtgc
     4441 cgggcaccag ggcgtagcgc ttcatggctt tgtgcagctg ctcgccggcg gtcccgtctt
     4501 cgagtgggta gaatggcgct gggcccttct taatgttttt ggcatcttcc atggtggctt
     4561 taccaacagt accggattgc caagcttgga acactgagaa gcgtgcaggc ggcgaccccg
     4621 actcgccctg ggcgcgggcg ggcgtggacg cgccgcggct gcggaggctg gggcaggggc
     4681 cggtcctgcg gcggcggaag ctggctgcgg cggcggatgg cggcggcggc tccccaagtt
     4741 ctgcgcgctg ggatctctcg cgactccccg gatccgcctc tatttgaagg gtctggaacg
     4801 gcacgggtcc gcctccgtcg tgacgtacat ggccatatat gggaagcagg aagccctaat
     4861 atggcaggac cggccgggag gacccggagt gacgtgaagc cccccatccg acagcgcccg
     4921 gcacctccat cctgcaccct agggcccgcg cccgcgcccg catcccaggc gcccaggctt
     4981 cggggaagcc tagagctcta gccgccgccg gggcctccgg gacccgcgca gctccggagt
     5041 tcccgcgttg cccctccctc cgccttcttc cctcctccca gagccggaag cggccgggcc
     5101 atatcgggcc actccaaata aggtgctgcc caaataaggg ttgttccggc gggggatcct
     5161 tcctgctcct tatatggcat ttccgggtcg cagctggttg ctccctcgct ccctccctcc
     5221 tccctggttc caagccggga gcccgagcgc tcgcgggatc cggggacgcc caggcgggga
     5281 gggctcgtgg cgtgtggcgc tgggcgcgcg gctgctcagg cggctcctag gcgtgaacta
     5341 ggctggggaa gcccagaccc agagcggggg cgagggcgag tcgggccggg gttctatcgc
     5401 tgtcatccag ggcccgcccg cccacctcag ggaccgtgag agcgaaccgg gagtgcgggg
     5461 gaggagcggc tgggcccgcc tgggcagggg ccgcccagga gaggcggggc cttgaccccg
     5521 ggcctcgatc tatggcacgg tgtctttccg ggtgtcgctg ggaaattgag gataggaagt
     5581 cacggccact tggttcttgg gacactgtag gtaccggcca gttaggccag agaaatgttc
     5641 tggcacctgc acttgcactg gggacagcct attttgctag tttgttttgt ttcgttttgt
     5701 tttgatggag agcgtatgtt agtactatcg attcacacaa aaaaccaaca cacagatgta
     5761 atgaaaataa agatatttta ttgcggcctg tccaatacct cccgtacctt aatattactt
     5821 acttatcctt gagagacgta ctagtaaccc tgataaatgc ttcaataata ttgaaaaagg
     5881 acgagtatga gcatccaaca ttttcgtgtc gcactcattc ccttctttgc ggcattttgc
     5941 ttgcctgttt ttgcacaccc cgaaacgctg gtgaaagtaa aagatgctga agatcaactg
     6001 ggtgcaagag tgggctatat cgaactggat ctcaatagcg gcaagatcct tgagtctttc
     6061 cgccccgaag aacgattccc gatgatgagc actttcaaag tactgctatg tggcgcggtg
     6121 ttgtcccgta tagacgccgg gcaagagcag cttggtcgcc gtatacacta ctcacaaaac
     6181 gacttggttg agtactcgcc ggtcacggaa aagcatctta cggatggcat gacggtaaga
     6241 gaattgtgta gtgctgccat taccatgagc gacaacaccg cggccaactt acttctgaca
     6301 acgatcggag gccctaagga gctgactgca tttcttcata atatgggtga tcatgtgacc
     6361 cggcttgacc gctgggaacc agagttgaac gaagccatac cgaacgacga gcgtgatacc
     6421 acgatgccag tagcaatggc cacaactctt cggaaactac tcactggcga acttcttact
     6481 ctagcatcac gacagcagct catagactgg atggaggcgg acaaagtagc aggaccactt
     6541 cttcgctcgg ccctccctgc tggctggttc attgctgaca aatcgggggc cggtgaacgc
     6601 ggctctcgcg gcatcattgc tgcgctgggg cctgatggta agccctcacg aatcgtagtg
     6661 atctacacga cggggagtca ggccactatg gacgaacgaa ataggcagat cgctgagatc
     6721 ggtgcctcac tgatcaagca ctggtaacca ctgcagtggt ttagcatttg cggccgctgt
     6781 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6841 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6901 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
