LOCUS       sequence-206333-        5903 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 5903)
  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 5903)
  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 1kb prom. CD274
FEATURES             Location/Qualifiers
     source          1..5903
                     /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      1334..1343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1340..2992
                     /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(1406..1424)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3033..3154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3044..3063)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3098..3117
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3402..3419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3573..4161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3653..3672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4332..5123,5124..5192))
                     /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     4955..4974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5193..5297)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5324..5779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5411..5430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5621..5642
                     /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 gagctcgaga tctgcgatct aagtaagctt ggcattccgg
     1321 tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt
     1381 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
     1441 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc
     1501 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
     1561 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
     1621 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
     1681 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca
     1741 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc
     1801 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
     1861 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact
     1921 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac
     1981 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
     2041 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     2101 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct
     2161 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt
     2221 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     2281 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc
     2341 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     2401 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     2461 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg
     2521 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     2581 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     2641 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt
     2701 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga
     2761 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     2821 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     2881 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     2941 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag
     3001 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3061 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3121 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3181 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3241 tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     3301 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     3361 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     3421 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3481 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3541 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3601 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3661 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3721 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3781 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3841 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3901 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3961 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4021 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4081 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4141 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4201 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4261 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4321 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4381 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4441 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4501 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4561 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4621 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4681 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4741 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4801 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4861 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4921 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4981 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5041 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5101 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5161 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5221 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5281 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     5341 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5401 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5461 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5521 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5581 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5641 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5701 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5761 atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5821 tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt aagtaatatt
     5881 aaggtacggg aggtacttgg agc
//
