LOCUS       sequence-479511-        5447 bp    DNA     circular SYN 23-JUL-2025
DEFINITION  Express IFNG response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5447)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5447)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 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            "Unpublished"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/IFNG/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..5447
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      709..718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             715..1227
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             1231..1350
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1402..1523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1413..1432)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1467..1486
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1717..2074
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1717..1908
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1740..1760)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1925..2060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1987..2006
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2105..3142
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3121..3129
                     /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    3166..3214
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3370..3387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3541..4129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3621..3640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4329..5120,5121..5189))
                     /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    5294..5342
                     /note="synthetic polyadenylation signal"
     misc_feature    5356..5447
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5396..5415
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gtgtggcttg tattgtattt ctactgggca gtgctgatct
       61 agagcaattt gaaacttgtg gtagatattt tactaaccaa ctctgatgaa ggacttcctc
      121 accaaattgt tcttttaacc gcattctttc cttgctttct ggtcatttgc aagaaaaatt
      181 ttaaaaggct gcccctttgt aaaggtttga gaggccctag aatttcgttt ttcacttgtt
      241 cccaaccaca agcaaatgat caatgtgctt tgtgaatgaa gagtcaacat tttaccaggg
      301 cgaagtgggg aggtacaaaa aaatttccag tccttgaatg gtgtgaagta aaagtgcctt
      361 caaagaatcc caccagaatg gcacaggtgg gcataatggg tctgtctcat cgtcaaagga
      421 cccaaggagt ctaaaggaaa ctctaactac aacacccaaa tgccacaaaa ccttagttat
      481 taatacaaac tatcatccct gcctatctgt caccatctca tcttaaaaaa cttgtgaaaa
      541 tacgtaatcc tcaggagact tcaattaggt ataaatacca gcagccagag gaggtgcagc
      601 acattgttct gatcatctga agatcagcta ttagaagaga aagatcagtt aagtcctttg
      661 gacctgatca gcttgataca agcttggcaa tccggtactg ttggtaaagc caccatggtc
      721 ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa
      781 gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg
      841 atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc
      901 ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg
      961 taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac
     1021 ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc
     1081 gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag
     1141 cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc
     1201 tggcggctgt gcgaacgcat tctggcgaat tctcacggct tccctcccga ggtggaggag
     1261 caggccgccg gcaccctgcc catgagctgc gcccaggaga gcggcatgga tagacaccct
     1321 gctgcttgcg ccagcgccag gatcaacgtc taaggccgcg actctagagt cggggcggcc
     1381 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1441 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1501 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1561 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     1621 ataaggatcc gtttgcgtat tgggcgctct tccgctgatc tgcgcagcac catggcctga
     1681 aataacctct gaaagaggaa cttggttagc taccttctga ggcggaaaga accagctgtg
     1741 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1801 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     1861 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     1921 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     1981 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2041 aggaggcttt tttggaggcc taggcttttg caaaaagctc gattcttctg acactagcgc
     2101 caccatgaag aagcccgaac tcaccgctac cagcgttgaa aaatttctca tcgagaagtt
     2161 cgacagtgtg agcgacctga tgcagttgtc ggagggcgaa gagagccgag ccttcagctt
     2221 cgatgtcggc ggacgcggct atgtactgcg ggtgaatagc tgcgctgatg gcttctacaa
     2281 agaccgctac gtgtaccgcc acttcgccag cgctgcacta cccatccccg aagtgttgga
     2341 catcggcgag ttcagcgaga gcctgacata ctgcatcagt agacgcgccc aaggcgttac
     2401 tctccaagac ctccccgaaa cagagctgcc tgctgtgtta cagcctgtcg ccgaagctat
     2461 ggatgctatt gccgccgccg acctcagtca aaccagcggc ttcggcccat tcgggcccca
     2521 aggcatcggc cagtacacaa cctggcggga tttcatttgc gccattgctg atccccatgt
     2581 ctaccactgg cagaccgtga tggacgacac cgtgtccgcc agcgtagctc aagccctgga
     2641 cgaactgatg ctgtgggccg aagactgtcc cgaggtgcgc cacctcgtcc atgccgactt
     2701 cggcagcaac aacgtcctga ccgacaacgg ccgcatcacc gccgtaatcg actggtccga
     2761 agctatgttc ggggacagtc agtacgaggt ggccaacatc ttcttctggc ggccctggct
     2821 ggcttgcatg gagcagcaga ctcgctactt cgagcgccgg catcccgagc tggccggcag
     2881 ccctcgtctg cgagcctaca tgctgcgcat cggcctggat cagctctacc agagcctcgt
     2941 ggacggcaac ttcgacgatg ctgcctgggc tcaaggccgc tgcgatgcca tcgtccgcag
     3001 cggggccggc accgtcggtc gcacacaaat cgctcgccgg agcgcagccg tatggaccga
     3061 cggctgcgtc gaggtgctgg ccgacagcgg caaccgccgg cccagtacac gaccgcgcgc
     3121 taaggaggta ggtcgagttt aaactctaga accggtcatg gccgcaataa aatatcttta
     3181 ttttcattac atctgtgtgt tggttttttg tgtgttcgaa ctagatgctg tcgaccgatg
     3241 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     3301 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3361 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3421 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3961 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4081 tttttttgtt tgcaagcagc agattacgca cagaaaaaaa ggatctcaag aagatccttt
     4141 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4201 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4261 atcaatctaa agtatatatg agtaaacttg gtctgacagc ggccgcaaat gctaaaccac
     4321 tgcagtggtt accagtgctt gatcagtgag gcaccgatct cagcgatctg cctatttcgt
     4381 tcgtccatag tggcctgact ccccgtcgtg tagatcacta cgattcgtga gggcttacca
     4441 tcaggcccca gcgcagcaat gatgccgcga gagccgcgtt caccggcccc cgatttgtca
     4501 gcaatgaacc agccagcagg gagggccgag cgaagaagtg gtcctgctac tttgtccgcc
     4561 tccatccagt ctatgagctg ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt
     4621 ttccgaagag ttgtggccat tgctactggc atcgtggtat cacgctcgtc gttcggtatg
     4681 gcttcgttca actctggttc ccagcggtca agccgggtca catgatcacc catattatga
     4741 agaaatgcag tcagctcctt agggcctccg atcgttgtca gaagtaagtt ggccgcggtg
     4801 ttgtcgctca tggtaatggc agcactacac aattctctta ccgtcatgcc atccgtaaga
     4861 tgcttttccg tgaccggcga gtactcaacc aagtcgtttt gtgagtagtg tatacggcga
     4921 ccaagctgct cttgcccggc gtctatacgg gacaacaccg cgccacatag cagtactttg
     4981 aaagtgctca tcatcgggaa tcgttcttcg gggcggaaag actcaaggat cttgccgcta
     5041 ttgagatcca gttcgatata gcccactctt gcacccagtt gatcttcagc atcttttact
     5101 ttcaccagcg tttcggggtg tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg
     5161 agtgcgacac gaaaatgttg gatgctcata ctcgtccttt ttcaatatta ttgaagcatt
     5221 tatcagggtt actagtacgt ctctcaagga taagtaagta atattaaggt acgggaggta
     5281 ttggacaggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     5341 tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc
     5401 aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctct
//
