LOCUS       sequence-472888-        6946 bp    DNA     circular SYN 14-MAY-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)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6946)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/hEGR1/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(20..377)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      34..169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(88..107)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        186..377
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     334..354
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    571..692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(608..627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     662..681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(744..863)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(867..2516)
                     /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      2513..2522
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3593..3684
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3625..3644)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3698..3746
                     /note="synthetic polyadenylation signal"
     CDS             join(3851..3919,3920..4711)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4911..5499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5400..5419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5653..5670
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5826..5874
                     /note="synthetic polyadenylation signal"
     CDS             complement(5898..6935)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5911..5919
                     /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)"
ORIGIN
        1 tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
       61 tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattagtc
      121 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
      181 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      241 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
      301 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
      361 ggttctttcc gcctcagaag gtagctaacc aagttcctct ttcagaggtt atttcaggcc
      421 atggtgctgc gcagatcagc ggaagagcgc ccaatacgca aacggatcct tatcgatttt
      481 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
      541 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
      601 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
      661 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gctcgaagcg gccggccgcc
      721 ccgactctag agtcgcggcc ttagacgttg atcctggcgc tggcgcaagc agcagggtgt
      781 ctatccatgc cgctctcctg ggcgcagctc atgggcaggg tgccggcggc ctgctcctcc
      841 acctcgggag ggaagccgtg agaattcacg gcgatcttgc cgcccttctt ggccttaatg
      901 agaatctcgc ggatcttgcg ggcgtccaac ttgccggtca gtcctttagg cacctcgtcc
      961 acgaacacaa caccaccgcg cagcttcttg gcggttgtaa cctggctggc cacatagtcc
     1021 acgatctcct tctcggtcat ggttttaccg tgttccagca cgacgactgc ggcgggcagc
     1081 tcgccggcat cgtcgtcggg caggccggcg accccggcgt cgaagatgtt ggggtgttgc
     1141 agcaggatgc tctccagttc ggctggggct acctggtagc ccttgtattt gatcaggctc
     1201 ttcagccggt ccacgatgaa gaagtgctcg tcctcgtccc agtaggcgat gtcgccgctg
     1261 tgcagccagc cgtccttgtc gatgagagcg tttgtagcct cggggttgtt aacgtagccg
     1321 ctcatgatca tggggccacg gacgcacagc tcgccgcgct ggttcacacc cagtgtctta
     1381 ccggtgtcca agtccaccac cttagcctcg aagaagggca ccaccttgcc tactgcgcca
     1441 ggcttgtcgt ccccttcggg ggtgatcaga atggcgctgg ttgtttctgt caggccgtag
     1501 ccctggcgga tgcctggtag gtggaagcgt ttggccacgg cctcacctac ctccttgctg
     1561 agcggcgccc cgccgctggc gatctcgtgc aagttgctta ggtcgtactt gtcgatgaga
     1621 gtgctcttag cgaagaagct aaatagtgtg ggcaccagca gggcagattg aatcttatag
     1681 tcttgcaagc tgcgcaagaa tagctcctcc tcgaagcggt acatgagcac gacccgaaag
     1741 ccgcagatca agtagcccag cgtggtgaac atgccgaagc cgtggtgaaa tggcaccacg
     1801 ctgaggatag cggtgtcggg gatgatctgg ttgccgaaga tggggtcgcg ggcatgactg
     1861 aatcggacac aagcggtgcg gtgcggtagg gctacgccct tgggcaatcc ggtactgcca
     1921 ctactgttca tgatcagggc gatggttttg tcccggtcga agctctcggg cacgaagtcg
     1981 tactcgttga agccgggtgg caaatgggaa gtcacgaagg tgtacatgct ttggaagccc
     2041 tggtagtcgg tcttgctatc catgatgatg atcttttgta tgatcggtag cttcttttgc
     2101 acgttgagga tcttttgcag ccctttcttg ctcacgaata cgacggtggg ctggctgatg
     2161 cccatgctgt tcagcagctc gcgctcgttg tagatgtcgt tagctggggc cacagccaca
     2221 ccgatgaaca gggcacccaa cacgggcatg aagaactgca agctattctc gctgcacacc
     2281 acgatccgat ggtttgtatt cagcccatag cgcttcatag cttctgccag ccgaacgctc
     2341 atctcgaagt actcggcgta ggtaatgtcc acctcgatat gtgcgtcggt aaaggcgatg
     2401 gtgccgggca ccagggcgta gcgcttcatg gctttgtgca gctgctcgcc ggcggtcccg
     2461 tcttcgagtg ggtagaatgg cgctgggccc ttcttaatgt ttttggcatc ttccatggtg
     2521 gctttaccaa cagtaccgga ttgccaagct tggaacactg agaagcgtgc aggcggcgac
     2581 cccgactcgc cctgggcgcg ggcgggcgtg gacgcgccgc ggctgcggag gctggggcag
     2641 gggccggtcc tgcggcggcg gaagctggct gcggcggcgg atggcggcgg cggctcccca
     2701 agttctgcgc gctgggatct ctcgcgactc cccggatccg cctctatttg aagggtctgg
     2761 aacggcacgg gtccgcctcc gtcgtgacgt acatggccat atatgggaag caggaagccc
     2821 taatatggca ggaccggccg ggaggacccg gagtgacgtg aagcccccca tccgacagcg
     2881 cccggcacct ccatcctgca ccctagggcc cgcgcccgcg cccgcatccc aggcgcccag
     2941 gcttcgggga agcctagagc tctagccgcc gccggggcct ccgggacccg cgcagctccg
     3001 gagttcccgc gttgcccctc cctccgcctt cttccctcct cccagagccg gaagcggccg
     3061 ggccatatcg ggccactcca aataaggtgc tgcccaaata agggttgttc cggcggggga
     3121 tccttcctgc tccttatatg gcatttccgg gtcgcagctg gttgctccct cgctccctcc
     3181 ctcctccctg gttccaagcc gggagcccga gcgctcgcgg gatccgggga cgcccaggcg
     3241 gggagggctc gtggcgtgtg gcgctgggcg cgcggctgct caggcggctc ctaggcgtga
     3301 actaggctgg ggaagcccag acccagagcg ggggcgaggg cgagtcgggc cggggttcta
     3361 tcgctgtcat ccagggcccg cccgcccacc tcagggaccg tgagagcgaa ccgggagtgc
     3421 gggggaggag cggctgggcc cgcctgggca ggggccgccc aggagaggcg gggccttgac
     3481 cccgggcctc gatctatggc acggtgtctt tccgggtgtc gctgggaaat tgaggatagg
     3541 aagtcacggc cacttggttc ttgggacact gtaggtaccg gccagttagg ccagagaaat
     3601 gttctggcac ctgcacttgc actggggaca gcctattttg ctagtttgtt ttgtttcgtt
     3661 ttgttttgat ggagagcgta tgttagtact atcgattcac acaaaaaacc aacacacaga
     3721 tgtaatgaaa ataaagatat tttattgcgg cctgtccaat acctcccgta ccttaatatt
     3781 acttacttat ccttgagaga cgtactagta accctgataa atgcttcaat aatattgaaa
     3841 aaggacgagt atgagcatcc aacattttcg tgtcgcactc attcccttct ttgcggcatt
     3901 ttgcttgcct gtttttgcac accccgaaac gctggtgaaa gtaaaagatg ctgaagatca
     3961 actgggtgca agagtgggct atatcgaact ggatctcaat agcggcaaga tccttgagtc
     4021 tttccgcccc gaagaacgat tcccgatgat gagcactttc aaagtactgc tatgtggcgc
     4081 ggtgttgtcc cgtatagacg ccgggcaaga gcagcttggt cgccgtatac actactcaca
     4141 aaacgacttg gttgagtact cgccggtcac ggaaaagcat cttacggatg gcatgacggt
     4201 aagagaattg tgtagtgctg ccattaccat gagcgacaac accgcggcca acttacttct
     4261 gacaacgatc ggaggcccta aggagctgac tgcatttctt cataatatgg gtgatcatgt
     4321 gacccggctt gaccgctggg aaccagagtt gaacgaagcc ataccgaacg acgagcgtga
     4381 taccacgatg ccagtagcaa tggccacaac tcttcggaaa ctactcactg gcgaacttct
     4441 tactctagca tcacgacagc agctcataga ctggatggag gcggacaaag tagcaggacc
     4501 acttcttcgc tcggccctcc ctgctggctg gttcattgct gacaaatcgg gggccggtga
     4561 acgcggctct cgcggcatca ttgctgcgct ggggcctgat ggtaagccct cacgaatcgt
     4621 agtgatctac acgacgggga gtcaggccac tatggacgaa cgaaataggc agatcgctga
     4681 gatcggtgcc tcactgatca agcactggta accactgcag tggtttagca tttgcggccg
     4741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5581 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5641 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgctgccg gcacctgtcc
     5701 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     5761 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac agcatctagt
     5821 tcgaacacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt tattgcggcc
     5881 atgaccggtt ctagagttta aactcgacct acctccttag cgcgcggtcg tgtactgggc
     5941 cggcggttgc cgctgtcggc cagcacctcg acgcagccgt cggtccatac ggctgcgctc
     6001 cggcgagcga tttgtgtgcg accgacggtg ccggccccgc tgcggacgat ggcatcgcag
     6061 cggccttgag cccaggcagc atcgtcgaag ttgccgtcca cgaggctctg gtagagctga
     6121 tccaggccga tgcgcagcat gtaggctcgc agacgagggc tgccggccag ctcgggatgc
     6181 cggcgctcga agtagcgagt ctgctgctcc atgcaagcca gccagggccg ccagaagaag
     6241 atgttggcca cctcgtactg actgtccccg aacatagctt cggaccagtc gattacggcg
     6301 gtgatgcggc cgttgtcggt caggacgttg ttgctgccga agtcggcatg gacgaggtgg
     6361 cgcacctcgg gacagtcttc ggcccacagc atcagttcgt ccagggcttg agctacgctg
     6421 gcggacacgg tgtcgtccat cacggtctgc cagtggtaga catggggatc agcaatggcg
     6481 caaatgaaat cccgccaggt tgtgtactgg ccgatgcctt ggggcccgaa tgggccgaag
     6541 ccgctggttt gactgaggtc ggcggcggca atagcatcca tagcttcggc gacaggctgt
     6601 aacacagcag gcagctctgt ttcggggagg tcttggagag taacgccttg ggcgcgtcta
     6661 ctgatgcagt atgtcaggct ctcgctgaac tcgccgatgt ccaacacttc ggggatgggt
     6721 agtgcagcgc tggcgaagtg gcggtacacg tagcggtctt tgtagaagcc atcagcgcag
     6781 ctattcaccc gcagtacata gccgcgtccg ccgacatcga agctgaaggc tcggctctct
     6841 tcgccctccg acaactgcat caggtcgctc acactgtcga acttctcgat gagaaatttt
     6901 tcaacgctgg tagcggtgag ttcgggcttc ttcatggtgg cgctag
//
