LOCUS       sequence-206337-        5941 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 5941)
  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 5941)
  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 and exon1 fragment CD274
FEATURES             Location/Qualifiers
     source          1..5941
                     /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      1372..1381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1378..3030
                     /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(1444..1462)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3071..3192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3082..3101)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3136..3155
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3440..3457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3611..4199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3691..3710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4370..5161,5162..5230))
                     /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     4993..5012
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5231..5335)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5362..5817
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5449..5468)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5659..5680
                     /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 tacgcgtgct ttaatcttcg aaactcttcc cggtgaaaat ctcatttaca agaaaactgg
      241 actgacatgt ttcactttct gtttcatttc tatacacagc tttattccta ggacaccaac
      301 actagatacc taaactgaaa gcttccgccg atttcaccga aggtcaggaa agtccaacgc
      361 ccggcaaact ggatttgctg ccttgggcag aggtgggcgg gaccccgcct ccgggcctgg
      421 cgcaacgctg agcagctggc gcgtcccgcg cggccccagt tctgcgcagc ttcccgaggc
      481 tccgcaccag ccgcgcttct gtccgcctgc aggtagggag cgttgttcct ccgcgggtgc
      541 ccacggccca gtatctctgg ctagctcgct gggcacttta ggacggaggg tctctacacc
      601 ctttctttgg gatggagaga ggagaaggga aagggaacgc gatggtctag ggggcagtag
      661 agccaattac ctgttggggt taataagaac aggcaatgca tctggccttc ctccaggcgc
      721 gattcagttt tgctctaaaa ataatttata cctctaaaaa taaataagat aggtagtata
      781 ggataggtag tcattcttat gcgactgtgt gttcagaata tagctctgat gctaggctgg
      841 aggtctggac acgggtccaa gtccaccgcc agctgcttgc tagtaacatg acttgtgtaa
      901 gttatcccag ctgcagcatc taagtaagtc tcttcctgcg ctaagcaggt ccaggatccc
      961 tgaacggaat ttatttgctc tgtccattct gagaacccaa aggagtccta aaagaggaat
     1021 ggaggagcct aagaataaaa atagtataat aaaacatttc ttagacacat tgaccttggc
     1081 ctatgtcaaa gttcagtctg ggtttgtctt ataacacaag gagtaaaagt accattgttc
     1141 tacctctttt tttaatactt gaaaaaaatt tactgtggat gcttttctat gaattaaata
     1201 accttctaaa aaatgttttc attgctgcat tcgattagat tgggtaacta aatgaaatta
     1261 attcctcact gttgggtata aaggttattt acagtggttc tgtcttagcc attcactgaa
     1321 cctcgagatc tgcgatctaa gtaagcttgg cattccggta ctgttggtaa agccaccatg
     1381 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
     1441 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
     1501 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
     1561 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
     1621 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
     1681 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
     1741 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
     1801 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
     1861 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     1921 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
     1981 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     2041 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     2101 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     2161 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     2221 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     2281 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     2341 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     2401 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     2461 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     2521 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     2581 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     2641 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     2701 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     2761 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     2821 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     2881 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     2941 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     3001 gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga
     3061 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3121 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3181 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3241 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     3301 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     3361 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     3421 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3481 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3541 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3601 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3661 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3721 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3781 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3841 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3901 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3961 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4021 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4081 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4141 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4201 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4261 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4321 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4381 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4441 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4501 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4561 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4621 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4681 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4741 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4801 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4861 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4921 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4981 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5041 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5101 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5161 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5221 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5281 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5341 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5401 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5461 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5521 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5581 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5641 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5701 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5761 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc
     5821 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5881 tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag
     5941 c
//
