LOCUS       sequence-206327-        6166 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 6166)
  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 6166)
  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'UTR reporter WT 1.3 kb CD274 Hs 3'UTR F...
FEATURES             Location/Qualifiers
     source          1..6166
                     /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"
     promoter        215..411
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      262..397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      441..450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             447..2099
                     /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(513..531)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             complement(2526..2537)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3665..3682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3836..4424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3916..3935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4595..5386,5387..5455))
                     /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     5218..5237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5456..5560)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5587..6042
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5674..5693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5884..5905
                     /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 agcccgggct cgagatctgc gatctgcatc tcaattagtc agcaaccata
      241 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      301 ccccatcgct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      361 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttggc
      421 attccggtac tgttggtaaa gccaccatgg aagacgccaa aaacataaag aaaggcccgg
      481 cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga
      541 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca
      601 cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc
      661 tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg
      721 tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac
      781 gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg
      841 ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca
      901 tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc
      961 tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa
     1021 ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc
     1081 atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca
     1141 ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta
     1201 cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc
     1261 tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat
     1321 tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa
     1381 ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc
     1441 atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta
     1501 cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga
     1561 aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg
     1621 tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga
     1681 ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact
     1741 tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg
     1801 ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc
     1861 ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga
     1921 cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt
     1981 tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg
     2041 caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat
     2101 tctagagaaa aataacactg gaattccttt tctagcatta tatttattcc tgatttgcct
     2161 ttgccatata atctaatgct tgtttatata gtgtctggta ttgtttaaca gttctgtctt
     2221 ttctatttaa atgccactaa attttaaatt catacctttc catgattcaa aattcaaaag
     2281 atcccatggg agatggttgg aaaatctcca cttcatcctc caagccattc aagtttcctt
     2341 tccagaagca actgctactg cctttcattc atatgttctt ctaaagatag tctacatttg
     2401 gaaatgtatg ttaaaagcac gtatttttaa aatttttttc ctaaatagta acacattgta
     2461 tgtctgctgt gtactttgct atttttattt attttagtgt ttcttatata gcagatggaa
     2521 tgaatttgaa gttcccaggg ctgaggatcc atgccttctt tgtttctaag ttatctttcc
     2581 catagctttt cattatcttt catatgatcc agtatatgtt aaatatgtcc tacatataca
     2641 tttagacaac caccatttgt taagtatttg ctctaggaca gagtttggat ttgtttatgt
     2701 ttgctcaaaa ggagacccat gggctctcca gggtgcactg agtcaatcta gtcctaaaaa
     2761 gcaatcttat tattaactct gtatgacaga atcatgtctg gaacttttgt tttctgcttt
     2821 ctgtcaagta taaacttcac tttgatgctg tacttgcaaa atcacatttt ctttctggaa
     2881 attccggcag tgtaccttga ctgctagcta ccctgtgcca gaaaagcctc attcgttgtg
     2941 cttgaaccct tgaatgccac cagctgtcat cactacacag ccctcctaag aggcttcctg
     3001 gaggtttcga gattcagatg ccctgggaga tcccagagtt tcctttccct cttggccata
     3061 ttctggtgtc aatgacaagg agtaccttgg ctttgccaca tgtcaaggct gaagaaacag
     3121 tgtctccaac agagctcctt gtgttatctg tttgtacatg tgcatttgta cagtaattgg
     3181 tgtgacagtg ttctttgtgt gaattacagg caagaattgt ggctgagcaa ggcacatagt
     3241 ctactcagtc tattcctaag tcctaactcc tccttgtggt gttggatttg taaggcactt
     3301 tatccctttt gtctcatgtt tcatcgtaaa tggcataggc agagatgata cctaattctg
     3361 catttgattg tcactttttg tacctgcatt aatttaataa aatattctta tttattttgt
     3421 tacttggtac accagcatgt ccattttctt gtttattttg tgtttaataa aatgttcagt
     3481 ttaacatccc agtggagaaa gttacttgga atatttgcgg atccgtcgac cgatgccctt
     3541 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     3601 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
     3661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4561 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4621 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4681 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4741 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4801 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4861 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4921 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4981 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5041 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5101 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5161 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5221 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5281 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5341 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5401 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5461 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5521 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5581 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5641 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5701 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5761 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5821 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5881 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5941 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6001 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc
     6061 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc
     6121 taccatgata agtaagtaat attaaggtac gggaggtact tggagc
//
