LOCUS       sequence-189306-        7037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express IKKalpha in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7037)
  AUTHORS   Marazioti A, Lilis I, Vreka M, Apostolopoulou H, Kalogeropoulou A, 
            Giopanou I, Giotopoulou GA, Krontira AC, Iliopoulou M, Kanellakis 
            NI, Agalioti T, Giannou AD, Jones-Paris C, Iwakura Y, Kardamakis D, 
            Blackwell TS, Taraviras S, Spella M, Stathopoulos GT
  TITLE     Myeloid-derived interleukin-1beta drives oncogenic KRAS-NF-kappaBeta
            addiction in malignant pleural effusion.
  JOURNAL   Nat Commun. 2018 Feb 14;9(1):672. doi: 10.1038/s41467-018-03051-z.
  PUBMED    29445180
REFERENCE   2  (bases 1 to 7037)
  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"; journalName: "Nat 
            Commun."; date: "2018-02-14"; volume: "9(1):672. doi"; pages: " 
            10.1038/s41467-018-03051-z"
COMMENT     Sequence Label: pChuk
FEATURES             Location/Qualifiers
     source          1..7037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        778..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1082..1285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1236..1256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1324..1333
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1330..1332,1333..1335,1336..2046)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1330..1332,1333..1335,1336..2046)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1375..1396)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1636..1655)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1983..2004
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             4076..4084
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      4375..4384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4379..4388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4542..4663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4579..4598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4633..4652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4670..5125)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4807..4828)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5019..5038
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5152..5256
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5258..5615
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5258..5449
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5281..5301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5466..5601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5650..6444
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5650..6444
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5704..5723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6314..6333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6632..6651)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6676..6723
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
       61 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      121 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
      181 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      241 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      301 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      361 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      421 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      481 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      541 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      601 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      661 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
      721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1261 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
     1321 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1381 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1441 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1501 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1561 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1621 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1681 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1741 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1801 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1861 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1921 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1981 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2041 tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cctgcaggcg
     2101 gccgcgaatt cactagtgat tatggagcgg cccccggggc tgcggccggg cgcgggcggc
     2161 ccctgggaga tgcgggaacg gcttggcacc ggcggtttcg ggaacgtcag tctgtaccag
     2221 caccgggaac ttgatctcaa aatagcaatt aagtcttgtc gtttagagct aagttccaaa
     2281 aacagagagc gatggtgcca tgaaatccag atcatgaaaa agttggacca tgcgaatgtt
     2341 gtaaaggcct gtgatgtccc tgaggaattg aactttttaa ttaacgatgt gcctcttctg
     2401 gcaatggagt actgttctgg aggggacctc cggaagctac tcaacaaacc agaaaattgt
     2461 tgtggactta aagaaagcca gatactttct ttactgagtg acataggatc tgggatccga
     2521 tatctgcatg aaaacaaaat tatacatcga gatctaaaac ctgaaaatat agttcttcaa
     2581 gatgttggtg ggaagacaat acataaaata attgatttgg gttatgccaa agatgttgat
     2641 caaggaagtc tctgtacatc ttttgtggga acattgcagt atttggcccc agagctcttt
     2701 gaaaataagc cgtacacagc cactgtggat tattggagct ttgggaccat ggtgtttgaa
     2761 tgtattgctg gatataggcc ttttttgcat catctgcagc catttacctg gcatgagaag
     2821 attaagaaga aagatccaaa gtgtatattt gcatgtgaag agatgactgg agaagttcgg
     2881 tttagtagcc atttacctca gccaaacagc ctttgtagtt taatagtaga gccaatggaa
     2941 agctggctcc aattgatgct gaattgggac ccacagcaga gagggggacc tattgatctt
     3001 actttgaagc agccaagatg ttttgcatta atggatcaca ttctcaattt aaagatagtg
     3061 cacatcctaa atatgacttc tgcaaaaatc atttcttttc tgttaccatg tgatgaaagt
     3121 cttcattcac tacagtctcg aattgagcgt gaaacaggaa taaatacagg ttctcaggag
     3181 cttctgtcag agacagggat ttctctggat cctcggaaac cagcctctca gtgtgttcta
     3241 gatggagtta gaggctgtga tagctacatg gtttatttgt ttgataaaag taagactgta
     3301 tatgaaggac catttgcatc cagaagttta tctgattgtg taaattatat tgtacaagac
     3361 agcaaaatac aactgccaat tatacagctg cggaaagtat gggctgaagc agtgcactac
     3421 gtatctgggc taaaggaaga ctacagcagg ctcttccagg gacaaagagc agcaatgtta
     3481 agtcttctta gatataatgc taacttgaca aaaatgaaga atactttgat ctcagcatca
     3541 cagcaactca aagctaaatt ggagtttttt cgaaaaagca ttcagcttga cttggagaga
     3601 tatagtgagc agatgactta tgggatatct tcagaaaaaa tgttaaaagc atggaaagaa
     3661 atggaagaaa aggccattca ctattctgag gttggtgtca ttggttatct tgaggatcaa
     3721 attatgtctt tgcacactga aatcatggag ctgcagaaga gcccctacgg acgacgccag
     3781 ggagacttga tggagtctct ggagcagcgt gccattgatc tctataagca gctaaagcac
     3841 agacctcctg atcacttgta cagcgacagc acagagatgg tgaagatcat cgtgcacacc
     3901 gtgcagagtc aggaccgtgt tctcaaggag ctgtttggtc acctgagcaa gttgttgggc
     3961 tgcaagcaga agattattga tctactcccc aaggtggaag tggccctcag taacatcaaa
     4021 gaagctgaca atactgtcat gtttatgcag ggaaagaggc agaaagaaat ttggcacctc
     4081 cttaaaattg cctgtacaca gagttctgcc cgctctcttg taggatccag tctagaaggc
     4141 acagtaaccc ctccagtatc agcatggctg ccccctacat tagcagaccg tgaacatcct
     4201 ctgacatgtg tggtaactcc tcaagatgga gagacgttag cacaaatgat agaagaaaat
     4261 ctgaactgtc ttggccattt aagtactatt attcgtgaag caaatgagga ccagagcagt
     4321 agtttgatga gtcttgattg gagttggtta gcagaatgaa atcgaattcc cgcggccgcc
     4381 atggcggccg ggagcatgcg acgtcgggcc cgggatccac cggatctaga taactgatca
     4441 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4501 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     4681 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4741 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4801 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4861 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4921 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4981 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5041 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5101 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5161 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5221 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     5281 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5341 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5401 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5461 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5521 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5581 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     5641 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5701 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5761 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5821 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5881 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5941 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6001 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6061 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6121 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     6181 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6241 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6301 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6361 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6421 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6481 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6541 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6601 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     6661 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     6721 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     6781 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     6841 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     6901 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6961 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7021 cagaccccgt agaaaag
//
