LOCUS       sequence-206328-        7879 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 7879)
  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 7879)
  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 3 kb prom. CD274
FEATURES             Location/Qualifiers
     source          1..7879
                     /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      3310..3319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3316..4968
                     /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(3382..3400)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5009..5130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5020..5039)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5074..5093
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5378..5395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5549..6137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5629..5648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6308..7099,7100..7168))
                     /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     6931..6950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7169..7273)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7300..7755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7387..7406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7597..7618
                     /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 tacgcgtctc cccagcttag gacaggaagg agagagacac agtgtgagcg agagaagcat
      241 ctgattgtga gtgtcagatg aggacagctg ctactctctg gtcacccata catgttaatt
      301 acccctgtca tttaaaccat tgttttgtgt aactagaaca aagtattctt tccacgaagt
      361 ctgtttgaca cgagacacat ataggatgtg agtctatgac aagggtaggg gcaacaaatg
      421 tgaggggttg gatggaaaca gaggaagaga aatgagcatt gcatagggca gtggtttaaa
      481 accctacatg atcaaaaata gaacctggag acaaatcttg atattgtctt tatacataat
      541 aaaatgcatt ttaaaatatt cgatgcaaac aaagcagttt gtcatgtctt ttgttttgag
      601 tctctttagt atatgctgct tttgaagatc aagaacattt actggaaatt gctccttcac
      661 caggaatttg ctcacatctc ttcaggtcca cttataagat cttgaaatca gtcctcctga
      721 gatcagtaca aacgaggatc atctctcctg ctttagaagc aaaagtggca agcagggtag
      781 acagaggcag aaggaaggat ggtactgata aatcagccaa attagatgga agaaagcgaa
      841 ttacagctgt acttgcaaag aactcttggc tcattttttc ccttaaatat taaatcacta
      901 gttttcaaaa cttcaaatgt gaacatgact cacctgagga caatgttaaa atgttgacct
      961 ccaagaccct gcccccaagt ttggatccca ggcagaaccc tggaatctgt attttaaaca
     1021 ttaaccccaa gtcgagtgta atgcaagtaa ccatcagaga cactctgaga aacagcacat
     1081 tggcaatgat gaatttatat gtctaaaaat gaatagagta gatgttacat gaataggaag
     1141 tggtggtatt caagatgacc atagtatcta gctcttcagc cctggctccc cactgctctt
     1201 ctcccatctc agcacttctc catctatttc gtccaaataa aaaaattcac acagagtttc
     1261 aggacttaac cccccactca ttaaccatct gttttgcttt acatattttt ctgaggtaat
     1321 aaaatttctc tttttctaaa cacagcctgt ttttcaatct ccgggtagtt gatcaattgt
     1381 atgggaaaat gaatggctga agggtagaaa caggtgggaa agatgaacaa aaacacgaat
     1441 cctcacatta ctaatacgca aatcactgag cagcaagctg agcaaatacc ctcaattccc
     1501 atcagcaact ttagagaaag gcaaattccg tttgcctcat tgatcattta ggtagaccct
     1561 gaacactgct ttcataaaac aaaaacaaaa tacccatccc cagtttaaaa aattattcat
     1621 agatcatcca ggccatctag gaggatatga tttaatcctg gctacttggt aaattatttg
     1681 cccaagttaa ctcagctagt tagtggtaga tggctctgaa gccagttgtt tttttttgtt
     1741 ttgttttttg cagacctcaa gagtcatgat gaactagcag atcataaagt ttatgccctg
     1801 ggtcttgacc atttttagaa aaataaaaca ttaaatgaaa atatcagagg gcattgcaga
     1861 tagtagatct aagtattttt tcatgaaact tgttgtacat gtgtgtgtca tacacagact
     1921 atatatatgc agtacctgta aactgtattg ccacataatg tctatatttt cctagaggtc
     1981 acagtcacca aagttgggaa gtcacccaac ttcgggaact ttgggaagtc acccaaactt
     2041 acagtcacca aaattgctct attctactat gtgacctcaa aagtgatttg aaagaaggaa
     2101 catctgagct gggcccaaac cctattgcaa ttttattggg gccaaagaga actccatgct
     2161 cctgccaaat caaggcagtg tcagcctcaa taatttccca gataaaaata aaaatctgtg
     2221 atacaatcag aatgtgaaaa ttcttatttt ggaagcaaat gtcataacca atgcaagggc
     2281 tatctcaata ttcattcatt atgcagtatt ttgaactgca gttgaaatga ataagaagga
     2341 aaggcaaaca acgaagagtc caatttctca atttagaaaa agagaaaaaa aagaaaaggg
     2401 agcacacagg cacggtggct caagcctgta atatcagcac tttggcggat cacttgaggt
     2461 caaggagttc gagaaaagag agcacctaga agttcagcgc gggataatac ttaagtaaat
     2521 tatgacacca tcgtctgtca tcttgggccc attcactaac ccaaagcttt caaaagggct
     2581 ttcttaaccc tcacctagaa taggcttccg cagccttaat ccttagggtg gcagaatatc
     2641 agggaccctg agcattctta aaagatgtag ctcgggatgg gaagttcttt taatgacaaa
     2701 gcaaatgaag tttcattatg tcgaggaact ttgaggaagt cacagaatcc acgatttaaa
     2761 aatatatttc ctattataca cccatacaca cacacacaca cctactttct agaataaaaa
     2821 ccaaagccat atgggtctgc tgctgacttt ttatatgttg tagagttata tcaagttatg
     2881 tcaagatgtt cagtcacctt gaagaggctt ttatcagaaa gggggacgcc tttctgataa
     2941 aggttaaggg gtaaccttaa gctcttaccc ctctgaaggt aaaatcaagg tgcgttcaga
     3001 tgttggcttg ttgtaaattt ctttttttat taataacata ctaaatgtgg atttgcttta
     3061 atcttcgaaa ctcttcccgg tgaaaatctc atttacaaga aaactggact gacatgtttc
     3121 actttctgtt tcatttctat acacagcttt attcctagga caccaacact agatacctaa
     3181 actgaaagct tccgccgatt tcaccgaagg tcaggaaagt ccaacgcccg gcaaactgga
     3241 tttgctgcct tgggcagagc tcgagatctg cgatctaagt aagcttggca ttccggtact
     3301 gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat
     3361 ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg
     3421 gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag
     3481 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat
     3541 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg
     3601 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc
     3661 aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa
     3721 attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa
     3781 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt
     3841 tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc
     3901 atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc
     3961 tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact
     4021 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat
     4081 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg
     4141 agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc
     4201 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt
     4261 ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt
     4321 atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg
     4381 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat
     4441 ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct
     4501 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat
     4561 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt
     4621 gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa
     4681 tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat
     4741 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa
     4801 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga
     4861 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc
     4921 agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg
     4981 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     5041 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     5101 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     5161 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     5221 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     5281 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     5341 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5401 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5461 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5521 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5581 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5641 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5701 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5761 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5821 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5881 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5941 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6001 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6061 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6121 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6181 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6241 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6301 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6361 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6421 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6481 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6541 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6601 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6661 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6721 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6781 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6841 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6901 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6961 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7021 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7081 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7141 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7201 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7261 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     7321 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     7381 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7441 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     7501 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7561 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7621 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     7681 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7741 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     7801 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     7861 tacgggaggt acttggagc
//
