LOCUS       sequence-206336-        7010 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 7010)
  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 7010)
  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. CD274
FEATURES             Location/Qualifiers
     source          1..7010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     291..312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    580..628
                     /note="synthetic polyadenylation signal"
     misc_feature    642..733
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     682..701
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      3014..3023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3020..4672
                     /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(3086..3104)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4713..4834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4724..4743)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4778..4797
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5082..5099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5253..5841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5333..5352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6012..6803,6804..6872))
                     /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     6635..6654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6873..6977)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(7004..7010,1..449)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
       61 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      121 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      181 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
      241 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      301 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      361 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      421 ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg
      481 ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt
      541 aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc tttattttca
      601 ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa
      661 aacaaaacga aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc
      721 agaacatttc tctatcgata ggtaccgagc tcttacgcgt gagacactct gagaaacagc
      781 acattggcaa tgatgaattt atatgtctaa aaatgaatag agtagatgtt acatgaatag
      841 gaagtggtgg tattcaagat gaccatagta tctagctctt cagccctggc tccccactgc
      901 tcttctccca tctcagcact tctccatcta tttcgtccaa ataaaaaaat tcacacagag
      961 tttcaggact taacccccca ctcattaacc atctgttttg ctttacatat ttttctgagg
     1021 taataaaatt tctctttttc taaacacagc ctgtttttca atctccgggt agttgatcaa
     1081 ttgtatggga aaatgaatgg ctgaagggta gaaacaggtg ggaaagatga acaaaaacac
     1141 gaatcctcac attactaata cgcaaatcac tgagcagcaa gctgagcaaa taccctcaat
     1201 tcccatcagc aactttagag aaaggcaaat tccgtttgcc tcattgatca tttaggtaga
     1261 ccctgaacac tgctttcata aaacaaaaac aaaataccca tccccagttt aaaaaattat
     1321 tcatagatca tccaggccat ctaggaggat atgatttaat cctggctact tggtaaatta
     1381 tttgcccaag ttaactcagc tagttagtgg tagatggctc tgaagccagt tgtttttttt
     1441 tgttttgttt tttgcagacc tcaagagtca tgatgaacta gcagatcata aagtttatgc
     1501 cctgggtctt gaccattttt agaaaaataa aacattaaat gaaaatatca gagggcattg
     1561 cagatagtag atctaagtat tttttcatga aacttgttgt acatgtgtgt gtcatacaca
     1621 gactatatat atgcagtacc tgtaaactgt attgccacat aatgtctata ttttcctaga
     1681 ggtcacagtc accaaagttg ggaagtcacc caacttcggg aactttggga agtcacccaa
     1741 acttacagtc accaaaattg ctctattcta ctatgtgacc tcaaaagtga tttgaaagaa
     1801 ggaacatctg agctgggccc aaaccctatt gcaattttat tggggccaaa gagaactcca
     1861 tgctcctgcc aaatcaaggc agtgtcagcc tcaataattt cccagataaa aataaaaatc
     1921 tgtgatacaa tcagaatgtg aaaattctta ttttggaagc aaatgtcata accaatgcaa
     1981 gggctatctc aatattcatt cattatgcag tattttgaac tgcagttgaa atgaataaga
     2041 aggaaaggca aacaacgaag agtccaattt ctcaatttag aaaaagagaa aaaaaagaaa
     2101 agggagcaca caggcacggt ggctcaagcc tgtaatatca gcactttggc ggatcacttg
     2161 aggtcaagga gttcgagaaa agagagcacc tagaagttca gcgcgggata atacttaagt
     2221 aaattatgac accatcgtct gtcatcttgg gcccattcac taacccaaag ctttcaaaag
     2281 ggctttctta accctcacct agaataggct tccgcagcct taatccttag ggtggcagaa
     2341 tatcagggac cctgagcatt cttaaaagat gtagctcggg atgggaagtt cttttaatga
     2401 caaagcaaat gaagtttcat tatgtcgagg aactttgagg aagtcacaga atccacgatt
     2461 taaaaatata tttcctatta tacacccata cacacacaca cacacctact ttctagaata
     2521 aaaaccaaag ccatatgggt ctgctgctga ctttttatat gttgtagagt tatatcaagt
     2581 tatgtcaaga tgttcagtca ccttgaagag gcttttatca gaaaggggga cgcctttctg
     2641 ataaaggtta aggggtaacc ttaagctctt acccctctga aggtaaaatc aaggtgcgtt
     2701 cagatgttgg cttgttgtaa atttcttttt ttattaataa catactaaat gtggatttgc
     2761 tttaatcttc gaaactcttc ccggtgaaaa tctcatttac aagaaaactg gactgacatg
     2821 tttcactttc tgtttcattt ctatacacag ctttattcct aggacaccaa cactagatac
     2881 ctaaactgaa agcttccgcc gatttcaccg aaggtcagga aagtccaacg cccggcaaac
     2941 tggatttgct gccttgggca gagctcgaga tctgcgatct aagtaagctt ggcattccgg
     3001 tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt
     3061 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
     3121 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc
     3181 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
     3241 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
     3301 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
     3361 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca
     3421 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc
     3481 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
     3541 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact
     3601 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac
     3661 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
     3721 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     3781 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct
     3841 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt
     3901 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     3961 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc
     4021 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     4081 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     4141 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg
     4201 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     4261 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     4321 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt
     4381 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga
     4441 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     4501 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     4561 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     4621 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag
     4681 tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     4741 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4801 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4861 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4921 tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     4981 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     5041 tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg
     5101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5161 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5221 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5281 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5341 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5401 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5461 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5521 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5581 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5641 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5701 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5761 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5821 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5881 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5941 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6001 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6061 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6121 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6181 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6241 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6301 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6361 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6421 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6481 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6541 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6601 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6661 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6721 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6781 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6841 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6901 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6961 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
//
