LOCUS       sequence-471824-        5996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express STAT4 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5996)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5996)
  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            "Unpublished"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/STAT4-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..5996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..94
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      121..130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             127..1776
                     /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             1780..1899
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1951..2072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1962..1981)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2016..2035
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2266..2623
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2266..2457
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2289..2309)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2474..2609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2536..2555
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2654..3691
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3670..3678
                     /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    3715..3763
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3919..3936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4090..4678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4170..4189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4878..5669,5670..5738))
                     /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    5843..5891
                     /note="synthetic polyadenylation signal"
     misc_feature    5905..5996
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5945..5964
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct acaacagcct gatttccccg aaatgacggc aagcttagac
       61 actagagggt atataatgga agctcgactt ccagcttggc aatccggtac tgttggtaaa
      121 gccaccatgg aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa
      181 gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc
      241 accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag
      301 atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc
      361 gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc
      421 ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg
      481 ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac
      541 gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac
      601 cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag
      661 tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt
      721 agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga
      781 ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc
      841 agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc
      901 ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa
      961 gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc
     1021 actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg
     1081 ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag
     1141 ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag
     1201 cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc
     1261 ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg
     1321 agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg
     1381 cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg
     1441 aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg
     1501 ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc
     1561 gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc
     1621 gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc
     1681 gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt
     1741 ctcattaagg ccaagaaggg cggcaagatc gccgtgaatt ctcacggctt ccctcccgag
     1801 gtggaggagc aggccgccgg caccctgccc atgagctgcg cccaggagag cggcatggat
     1861 agacaccctg ctgcttgcgc cagcgccagg atcaacgtct aaggccgcga ctctagagtc
     1921 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     1981 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2041 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2101 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2161 gtaaaatcga taaggatccg tttgcgtatt gggcgctctt ccgctgatct gcgcagcacc
     2221 atggcctgaa ataacctctg aaagaggaac ttggttagct accttctgag gcggaaagaa
     2281 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2341 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2401 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2461 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2521 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2581 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcg attcttctga
     2641 cactagcgcc accatgaaga agcccgaact caccgctacc agcgttgaaa aatttctcat
     2701 cgagaagttc gacagtgtga gcgacctgat gcagttgtcg gagggcgaag agagccgagc
     2761 cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg
     2821 cttctacaaa gaccgctacg tgtaccgcca cttcgccagc gctgcactac ccatccccga
     2881 agtgttggac atcggcgagt tcagcgagag cctgacatac tgcatcagta gacgcgccca
     2941 aggcgttact ctccaagacc tccccgaaac agagctgcct gctgtgttac agcctgtcgc
     3001 cgaagctatg gatgctattg ccgccgccga cctcagtcaa accagcggct tcggcccatt
     3061 cgggccccaa ggcatcggcc agtacacaac ctggcgggat ttcatttgcg ccattgctga
     3121 tccccatgtc taccactggc agaccgtgat ggacgacacc gtgtccgcca gcgtagctca
     3181 agccctggac gaactgatgc tgtgggccga agactgtccc gaggtgcgcc acctcgtcca
     3241 tgccgacttc ggcagcaaca acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga
     3301 ctggtccgaa gctatgttcg gggacagtca gtacgaggtg gccaacatct tcttctggcg
     3361 gccctggctg gcttgcatgg agcagcagac tcgctacttc gagcgccggc atcccgagct
     3421 ggccggcagc cctcgtctgc gagcctacat gctgcgcatc ggcctggatc agctctacca
     3481 gagcctcgtg gacggcaact tcgacgatgc tgcctgggct caaggccgct gcgatgccat
     3541 cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt
     3601 atggaccgac ggctgcgtcg aggtgctggc cgacagcggc aaccgccggc ccagtacacg
     3661 accgcgcgct aaggaggtag gtcgagttta aactctagaa ccggtcatgg ccgcaataaa
     3721 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgttcgaac tagatgctgt
     3781 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     3841 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     3901 gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4501 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagcg gccgcaaatg
     4861 ctaaaccact gcagtggtta ccagtgcttg atcagtgagg caccgatctc agcgatctgc
     4921 ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt agatcactac gattcgtgag
     4981 ggcttaccat caggccccag cgcagcaatg atgccgcgag agccgcgttc accggccccc
     5041 gatttgtcag caatgaacca gccagcaggg agggccgagc gaagaagtgg tcctgctact
     5101 ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca
     5161 gtgagtagtt tccgaagagt tgtggccatt gctactggca tcgtggtatc acgctcgtcg
     5221 ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac atgatcaccc
     5281 atattatgaa gaaatgcagt cagctcctta gggcctccga tcgttgtcag aagtaagttg
     5341 gccgcggtgt tgtcgctcat ggtaatggca gcactacaca attctcttac cgtcatgcca
     5401 tccgtaagat gcttttccgt gaccggcgag tactcaacca agtcgttttg tgagtagtgt
     5461 atacggcgac caagctgctc ttgcccggcg tctatacggg acaacaccgc gccacatagc
     5521 agtactttga aagtgctcat catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc
     5581 ttgccgctat tgagatccag ttcgatatag cccactcttg cacccagttg atcttcagca
     5641 tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag
     5701 aagggaatga gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt tcaatattat
     5761 tgaagcattt atcagggtta ctagtacgtc tctcaaggat aagtaagtaa tattaaggta
     5821 cgggaggtat tggacaggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     5881 ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa
     5941 caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctct
//
