LOCUS       sequence-206331-        6832 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 6832)
  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 6832)
  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 2 kb prom. and exon 1 fragment CD274
FEATURES             Location/Qualifiers
     source          1..6832
                     /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      2263..2272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2269..3921
                     /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(2335..2353)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3962..4083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3973..3992)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4027..4046
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4331..4348)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4502..5090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4582..4601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5261..6052,6053..6121))
                     /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     5884..5903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6122..6226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6253..6708
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6340..6359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6550..6571
                     /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 gaggtgggcg ggaccccgcc tccgggcctg gcgcaacgct
     1321 gagcagctgg cgcgtcccgc gcggccccag ttctgcgcag cttcccgagg ctccgcacca
     1381 gccgcgcttc tgtccgcctg caggtaggga gcgttgttcc tccgcgggtg cccacggccc
     1441 agtatctctg gctagctcgc tgggcacttt aggacggagg gtctctacac cctttctttg
     1501 ggatggagag aggagaaggg aaagggaacg cgatggtcta gggggcagta gagccaatta
     1561 cctgttgggg ttaataagaa caggcaatgc atctggcctt cctccaggcg cgattcagtt
     1621 ttgctctaaa aataatttat acctctaaaa ataaataaga taggtagtat aggataggta
     1681 gtcattctta tgcgactgtg tgttcagaat atagctctga tgctaggctg gaggtctgga
     1741 cacgggtcca agtccaccgc cagctgcttg ctagtaacat gacttgtgta agttatccca
     1801 gctgcagcat ctaagtaagt ctcttcctgc gctaagcagg tccaggatcc ctgaacggaa
     1861 tttatttgct ctgtccattc tgagaaccca aaggagtcct aaaagaggaa tggaggagcc
     1921 taagaataaa aatagtataa taaaacattt cttagacaca ttgaccttgg cctatgtcaa
     1981 agttcagtct gggtttgtct tataacacaa ggagtaaaag taccattgtt ctacctcttt
     2041 ttttaatact tgaaaaaaat ttactgtgga tgcttttcta tgaattaaat aaccttctaa
     2101 aaaatgtttt cattgctgca ttcgattaga ttgggtaact aaatgaaatt aattcctcac
     2161 tgttgggtat aaaggttatt tacagtggtt ctgtcttagc cattcactga acctcgagat
     2221 ctgcgatcta agtaagcttg gcattccggt actgttggta aagccaccat ggaagacgcc
     2281 aaaaacataa agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag
     2341 caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat
     2401 gcacatatcg aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca
     2461 gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac
     2521 tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc
     2581 gcgaacgaca tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc
     2641 gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca
     2701 atcatccaaa aaattattat catggattct aaaacggatt accagggatt tcagtcgatg
     2761 tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag
     2821 tccttcgata gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg
     2881 cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat
     2941 cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat
     3001 cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta
     3061 atgtatagat ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt
     3121 gcgctgctgg tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac
     3181 gatttatcta atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg
     3241 gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag
     3301 actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa
     3361 gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt
     3421 aatcaaagag gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat
     3481 ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct
     3541 tactgggacg aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac
     3601 aaaggctatc aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc
     3661 ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt
     3721 gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt
     3781 caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa
     3841 ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag
     3901 ggcggaaaga tcgccgtgta attctagagt cggggcggcc ggccgcttcg agcagacatg
     3961 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4021 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4081 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     4141 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat
     4201 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     4261 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     4321 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4381 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4441 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4681 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4921 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6241 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6421 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6601 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6661 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat
     6721 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6781 ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga gc
//
