LOCUS       sequence-206329-        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 MUT 1.3 kb CD274 Hs 3'UTR ...
FEATURES             Location/Qualifiers
     source          1..6166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    123..171
                     /note="synthetic polyadenylation signal"
     misc_feature    185..276
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     225..244
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        331..527
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      378..513
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      557..566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             563..2215
                     /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(629..647)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             complement(2642..2653)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3781..3798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4032..4051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4711..5502,5503..5571))
                     /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     5334..5353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5572..5676)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5703..6158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5790..5809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6000..6021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
       61 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
      121 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
      181 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
      241 tccccagtgc aagtgcaggt gccagaacat ttctctatcg ataggtaccg agctcttacg
      301 cgtgctagcc cgggctcgag atctgcgatc tgcatctcaa ttagtcagca accatagtcc
      361 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
      421 atcgctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
      481 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttggcattc
      541 cggtactgtt ggtaaagcca ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc
      601 attctatccg ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata
      661 cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta
      721 cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa
      781 tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt
      841 gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga
      901 attgctcaac agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt
      961 gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga
     1021 ttctaaaacg gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc
     1081 tcccggtttt aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc
     1141 actgatcatg aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag
     1201 aactgcctgc gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc
     1261 ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact
     1321 cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt
     1381 tctgaggagc cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc
     1441 cttcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc
     1501 ttctggtggc gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct
     1561 gccaggtatc aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc
     1621 cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt
     1681 tgtggatctg gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag
     1741 aggtcctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga
     1801 caaggatgga tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt
     1861 catcgttgac cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga
     1921 attggaatcc atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc
     1981 cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat
     2041 gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg
     2101 cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag
     2161 aaaaatcaga gagatcctca taaaggccaa gaagggcgga aagatcgccg tgtaattcta
     2221 gagaaaaata acactggaat tccttttcta gcattatatg tattcctgat ttgcctttgc
     2281 catataatct aatgcttgtt tatatagtgt ctggtattgt ttaacagttc tgtcttttct
     2341 atgtaaatgc cactaaattt taaattcata cctttccatg attcaaaatt caaaagatcc
     2401 catgggagat ggttggaaaa tctccacttc atcctccaag ccattcaagt ttcctttcca
     2461 gaagcaactg ctactgcctt tcattcatat gttcttctaa agatagtcta catttggaaa
     2521 tgtatgttaa aagcacgtat ttttaaaatt tttttcctaa atagtaacac attgtatgtc
     2581 tgctgtgtac tttgctattt ttatgtattt tagtgtttct tatatagcag atggaatgaa
     2641 tttgaagttc ccagggctga ggatccatgc cttctttgtt tctaagttat ctttcccata
     2701 gcttttcatt atctttcata tgatccagta tatgttaaat atgtcctaca tatacatgta
     2761 gacaaccacc atttgttaag tatttgctct aggacagagt ttggatttgt ttatgtttgc
     2821 tcaaaaggag acccatgggc tctccagggt gcactgagtc aatctagtcc taaaaagcaa
     2881 tcttattatt aactctgtat gacagaatca tgtctggaac ttttgttttc tgctttctgt
     2941 caagtataaa cttcactttg atgctgtact tgcaaaatca cattttcttt ctggaaattc
     3001 cggcagtgta ccttgactgc tagctaccct gtgccagaaa agcctcattc gttgtgcttg
     3061 aacccttgaa tgccaccagc tgtcatcact acacagccct cctaagaggc ttcctggagg
     3121 tttcgagatt cagatgccct gggagatccc agagtttcct ttccctcttg gccatattct
     3181 ggtgtcaatg acaaggagta ccttggcttt gccacatgtc aaggctgaag aaacagtgtc
     3241 tccaacagag ctccttgtgt tatctgtttg tacatgtgca tttgtacagt aattggtgtg
     3301 acagtgttct ttgtgtgaat tacaggcaag aattgtggct gagcaaggca catagtctac
     3361 tcagtctatt cctaagtcct aactcctcct tgtggtgttg gatttgtaag gcactttatc
     3421 ccttttgtct catgtttcat cgtaaatggc ataggcagag atgataccta attctgcatt
     3481 tgattgtcac tttttgtacc tgcattaatg taataaaata ttcttatgta ttttgttact
     3541 tggtacacca gcatgtccat tttcttgttt attttgtgtt taataaaatg ttcagtttaa
     3601 catcccagtg gagaaagtta cttggaatat ttgcggatcc gtcgaccgat gcccttgaga
     3661 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     3721 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     3781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4801 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4861 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4921 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5221 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5581 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5641 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5701 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5761 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5821 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5881 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5941 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6001 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     6061 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     6121 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcg
//
