LOCUS       sequence-206332-        6621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified Luciferase expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6621)
  AUTHORS   Coelho MA, de Carne Trecesson S, Rana S, Zecchin D, Moore C, 
            Molina-Arcas M, East P, Spencer-Dene B, Nye E, Barnouin K, Snijders 
            AP, Lai WS, Blackshear PJ, Downward J
  TITLE     Oncogenic RAS Signaling Promotes Tumor Immunoresistance by 
            Stabilizing PD-L1 mRNA.
  JOURNAL   Immunity. 2017 Dec 19;47(6):1083-1099.e6. doi: 
            10.1016/j.immuni.2017.11.016. Epub 2017 Dec 12.
  PUBMED    29246442
REFERENCE   2  (bases 1 to 6621)
  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"; doi: 
            "10.1016/j.immuni.2017.11.016"; journalName: "Immunity"; date: 
            "2017-12-19- 19"; volume: "47"; issue: "6"; pages: "1083-1099.e6"
COMMENT     Sequence Label: pGL3 1 kb prom. + Enhancer CD274
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    7..55
                     /note="synthetic polyadenylation signal"
     misc_feature    69..160
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     109..128
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      1334..1343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1340..2992
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1406..1424)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3033..3154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3044..3063)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3098..3117
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4120..4137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4291..4879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4371..4390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5050..5841,5842..5910))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5673..5692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5911..6015)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6042..6497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6129..6148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6339..6360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
       61 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
      121 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accgagctct
      181 tacgcgtgcc aaatcaaggc agtgtcagcc tcaataattt cccagataaa aataaaaatc
      241 tgtgatacaa tcagaatgtg aaaattctta ttttggaagc aaatgtcata accaatgcaa
      301 gggctatctc aatattcatt cattatgcag tattttgaac tgcagttgaa atgaataaga
      361 aggaaaggca aacaacgaag agtccaattt ctcaatttag aaaaagagaa aaaaaagaaa
      421 agggagcaca caggcacggt ggctcaagcc tgtaatatca gcactttggc ggatcacttg
      481 aggtcaagga gttcgagaaa agagagcacc tagaagttca gcgcgggata atacttaagt
      541 aaattatgac accatcgtct gtcatcttgg gcccattcac taacccaaag ctttcaaaag
      601 ggctttctta accctcacct agaataggct tccgcagcct taatccttag ggtggcagaa
      661 tatcagggac cctgagcatt cttaaaagat gtagctcggg atgggaagtt cttttaatga
      721 caaagcaaat gaagtttcat tatgtcgagg aactttgagg aagtcacaga atccacgatt
      781 taaaaatata tttcctatta tacacccata cacacacaca cacacctact ttctagaata
      841 aaaaccaaag ccatatgggt ctgctgctga ctttttatat gttgtagagt tatatcaagt
      901 tatgtcaaga tgttcagtca ccttgaagag gcttttatca gaaaggggga cgcctttctg
      961 ataaaggtta aggggtaacc ttaagctctt acccctctga aggtaaaatc aaggtgcgtt
     1021 cagatgttgg cttgttgtaa atttcttttt ttattaataa catactaaat gtggatttgc
     1081 tttaatcttc gaaactcttc ccggtgaaaa tctcatttac aagaaaactg gactgacatg
     1141 tttcactttc tgtttcattt ctatacacag ctttattcct aggacaccaa cactagatac
     1201 ctaaactgaa agcttccgcc gatttcaccg aaggtcagga aagtccaacg cccggcaaac
     1261 tggatttgct gccttgggca gagctcgaga tctgcgatct aagtaagctt ggcattccgg
     1321 tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt
     1381 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
     1441 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc
     1501 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
     1561 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
     1621 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
     1681 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca
     1741 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc
     1801 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
     1861 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact
     1921 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac
     1981 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
     2041 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     2101 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct
     2161 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt
     2221 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     2281 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc
     2341 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     2401 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     2461 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg
     2521 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     2581 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     2641 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt
     2701 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga
     2761 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     2821 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     2881 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     2941 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag
     3001 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3061 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3121 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3181 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3241 tggtaaaatc gataaggatc ccaaagattg gagaggcact aagagggaaa acttacctct
     3301 ttcttatcaa agtttgtaaa tatatgtata acagaagagg gagaaaatat taataaatgc
     3361 acagattggc tgaaatagag tataaatctt ttactcccct acttcaacat aaactgcaaa
     3421 aggagagtga cttttctttc actctgactt ccgtattcct catgcttaaa atagtgccta
     3481 gcacagaaga ggtgctcaat cagtgtttgc taaacgaaat aattagtcac atttcaagca
     3541 ggatgactaa atgaagaata gaatctaggc agatactctg gaagagtggc tgtgagtcat
     3601 tcatatctta gtatgaatta gtcaaatcca actctctccc cttcccactc cccactgtta
     3661 gtagaagaat ctgtttattg agagaataga tttataattt agaataagtg agaggggcag
     3721 aagaggagat tttgaaggat ggcacctgaa ggaggactag catggctgag acagtgaagt
     3781 ggaagtcttg aatagctaaa gggtaagatg aaagtattta gctgtagggg gaaaaagcat
     3841 tgacaggttg gaaaagtaaa agtcagattc tccttgctct gaaatgttgt acagggcagg
     3901 ttctactagg tatgttacaa tgcagaaaaa acatgaaata attgagagga atttggtgca
     3961 atattatctt cttggcttcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     4021 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     4081 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     4141 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4201 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4261 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4321 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4381 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4441 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4501 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4561 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4621 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4681 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4741 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4801 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4861 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4921 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4981 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5041 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5101 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5161 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5221 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5281 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5341 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5401 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5461 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5521 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5581 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5641 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5701 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5761 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5821 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5881 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5941 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6001 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     6061 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6121 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6181 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6241 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6301 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6361 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     6421 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6481 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6541 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     6601 ggtacgggag gtacttggag c
//
