LOCUS       sequence-473396-        4891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express STAT6 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4891)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4891)
  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            "Unpublished"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: pNL [NlucP/STAT6-RE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        95..126
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      153..162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             159..671
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             675..794
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    846..967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(857..876)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     911..930
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1161..1518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1161..1352
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1184..1204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1369..1504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1431..1450
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1549..2586
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2565..2573
                     /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    2610..2658
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2814..2831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2985..3573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3065..3084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3773..4564,4565..4633))
                     /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    4738..4786
                     /note="synthetic polyadenylation signal"
     misc_feature    4800..4891
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4840..4859
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacca gtttccgagg aactgggccc agtttccgag gaactgggcc
       61 cagtttccga ggaactgggc cgaagcttag acactagagg gtatataatg gaagctcgac
      121 ttccagcttg gcaatccggt actgttggta aagccaccat ggtcttcaca ctcgaagatt
      181 tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag
      241 gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc
      301 tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
      361 gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc
      421 atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
      481 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
      541 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
      601 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
      661 gcattctggc gaattctcac ggctttccgc ctgaggttga agagcaagcc gccggtacat
      721 tgcctatgtc ctgcgcacaa gaaagcggta tggaccggca cccagccgct tgtgcttcag
      781 ctcgcatcaa cgtctaaggc cgcgactcta gagtcggggc ggccggccgc ttcgagcaga
      841 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      901 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      961 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1021 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtttgc
     1081 gtattgggcg ctcttccgct gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     1141 ggaacttggt tagctacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     1201 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1261 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1321 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1381 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1441 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     1501 ggcctaggct tttgcaaaaa gctcgattct tctgacacta gcgccaccat gaagaagccc
     1561 gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac
     1621 ctgatgcagt tgtcggaggg cgaagagagc cgagccttca gcttcgatgt cggcggacgc
     1681 ggctatgtac tgcgggtgaa tagctgcgct gatggcttct acaaagaccg ctacgtgtac
     1741 cgccacttcg ccagcgctgc actacccatc cccgaagtgt tggacatcgg cgagttcagc
     1801 gagagcctga catactgcat cagtagacgc gcccaaggcg ttactctcca agacctcccc
     1861 gaaacagagc tgcctgctgt gttacagcct gtcgccgaag ctatggatgc tattgccgcc
     1921 gccgacctca gtcaaaccag cggcttcggc ccattcgggc cccaaggcat cggccagtac
     1981 acaacctggc gggatttcat ttgcgccatt gctgatcccc atgtctacca ctggcagacc
     2041 gtgatggacg acaccgtgtc cgccagcgta gctcaagccc tggacgaact gatgctgtgg
     2101 gccgaagact gtcccgaggt gcgccacctc gtccatgccg acttcggcag caacaacgtc
     2161 ctgaccgaca acggccgcat caccgccgta atcgactggt ccgaagctat gttcggggac
     2221 agtcagtacg aggtggccaa catcttcttc tggcggccct ggctggcttg catggagcag
     2281 cagactcgct acttcgagcg ccggcatccc gagctggccg gcagccctcg tctgcgagcc
     2341 tacatgctgc gcatcggcct ggatcagctc taccagagcc tcgtggacgg caacttcgac
     2401 gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc
     2461 ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg
     2521 ctggccgaca gcggcaaccg ccggcccagt acacgaccgc gcgctaagga ggtaggtcga
     2581 gtttaaactc tagaaccggt catggccgca ataaaatatc tttattttca ttacatctgt
     2641 gtgttggttt tttgtgtgtt cgaactagat gctgtcgacc gatgcccttg agagccttca
     2701 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     2761 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
     2821 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2881 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2941 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3001 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3061 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3121 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3181 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3241 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3301 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3361 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3421 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3481 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3541 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3601 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3661 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3721 tatgagtaaa cttggtctga cagcggccgc aaatgctaaa ccactgcagt ggttaccagt
     3781 gcttgatcag tgaggcaccg atctcagcga tctgcctatt tcgttcgtcc atagtggcct
     3841 gactccccgt cgtgtagatc actacgattc gtgagggctt accatcaggc cccagcgcag
     3901 caatgatgcc gcgagagccg cgttcaccgg cccccgattt gtcagcaatg aaccagccag
     3961 cagggagggc cgagcgaaga agtggtcctg ctactttgtc cgcctccatc cagtctatga
     4021 gctgctgtcg tgatgctaga gtaagaagtt cgccagtgag tagtttccga agagttgtgg
     4081 ccattgctac tggcatcgtg gtatcacgct cgtcgttcgg tatggcttcg ttcaactctg
     4141 gttcccagcg gtcaagccgg gtcacatgat cacccatatt atgaagaaat gcagtcagct
     4201 ccttagggcc tccgatcgtt gtcagaagta agttggccgc ggtgttgtcg ctcatggtaa
     4261 tggcagcact acacaattct cttaccgtca tgccatccgt aagatgcttt tccgtgaccg
     4321 gcgagtactc aaccaagtcg ttttgtgagt agtgtatacg gcgaccaagc tgctcttgcc
     4381 cggcgtctat acgggacaac accgcgccac atagcagtac tttgaaagtg ctcatcatcg
     4441 ggaatcgttc ttcggggcgg aaagactcaa ggatcttgcc gctattgaga tccagttcga
     4501 tatagcccac tcttgcaccc agttgatctt cagcatcttt tactttcacc agcgtttcgg
     4561 ggtgtgcaaa aacaggcaag caaaatgccg caaagaaggg aatgagtgcg acacgaaaat
     4621 gttggatgct catactcgtc ctttttcaat attattgaag catttatcag ggttactagt
     4681 acgtctctca aggataagta agtaatatta aggtacggga ggtattggac aggccgcaat
     4741 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta
     4801 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
     4861 agtgcaagtg caggtgccag aacatttctc t
//
