LOCUS       sequence-140320-        7039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse eGFP-IKK-beta in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7039)
  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 7039)
  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: pEGFP.mIKK-beta
FEATURES             Location/Qualifiers
     source          1..7039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1406..1415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1410..1419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3827..3948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3864..3883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3918..3937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3955..4410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4092..4113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4304..4323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4437..4541
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4543..4900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4543..4734
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4566..4586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4751..4886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4935..5729
                     /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             4935..5729
                     /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(4989..5008)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5599..5618
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5917..5936)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5961..6008
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6337..6925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6826..6845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 gcgggcccga cgtcgcatgc tcccggccgc catggcggcc gcgggaattc gattatgagc
     1441 tggtcaccgt ccctcccaac ccagacatgt ggagcctggg aaatgaaaga acgcctgggg
     1501 accgggggat ttggaaacgt catccggtgg cacaatcagg cgacaggtga acagatcgcc
     1561 atcaagcaat gccgacagga gctcagccca aagaacagag accgctggtg cctcgaaatc
     1621 cagatcatga gaaggctgaa ccatcccaat gtggtggctg cccgggatgt cccagagggg
     1681 atgcagaacc tggcacccaa tgatttgccc ctgctggcca tggagtactg ccaaggagga
     1741 gatctccgaa gatacttgaa ccagttcgag aactgctgtg gcctgcggga aggagctgtc
     1801 cttaccctgt tgagtgacat cgcatcggct cttggatacc ttcacgaaaa cagaatcatc
     1861 catcgagacc tgaagccaga aaacatcgtt ctgcagcaag gagagaaaag attaatacac
     1921 aaaattattg atctaggata tgccaaggag ctggatcagg gcagtctgtg cacgtcattt
     1981 gtggggactc tgcaatacct ggcgccagag cttctggagc agcagaagta caccgtgacc
     2041 gttgactact ggagcttcgg caccctggcc ttcgagtgca tcactggctt ccggcccttc
     2101 ctccctaact ggcagcctgt gcagtggcac tccaaagtcc ggcagaagag cgaagtggac
     2161 atcgttgtta gtgaagactt gaatggagca gtgaagtttt caagttcgct acccttcccc
     2221 aataatctta acagtgtctt ggctgaacgg ctggagaagt ggctgcagct gatgcttatg
     2281 tggcaccctc ggcaaagggg cacggatccc cagtatggcc ccaacggctg cttcagagcc
     2341 ctggatgaca tcttgaactt gaagctggtt catgtcttga acatggtcac aggcaccgtt
     2401 cacacatacc ccgtgacgga ggatgagagt ctgcagagct taaaaaccag aatccaggaa
     2461 gacacgggga tcctggagac agaccaggag ctgctgcaag aggcagggct ggtgctgctc
     2521 cctgacaagc ctgctactca gtgcatctca gacagcaaga caaacgaggg cctcacgttg
     2581 gacatggatc ttgtttttct ctttgacaac agtaaaatca actatgagac tcagatcacc
     2641 ccccgacccc aaccggaaag tgtcagctgt atccttcagg agcccaagcg gaacctctcc
     2701 ttcttccagc tgaggaaagt gtggggccaa gtctggcaca gcatccagac gctgaaggaa
     2761 gactgtaacc ggctgcagca gggacagcga gcagccatga tgagtctcct ccggaataac
     2821 agctgcctct ctaagatgaa gaacgccatg gcctccacgg cccagcagct caaggccaag
     2881 ctggacttct tcaaaaccag catccagatc gacctggaga agtataaaga gcagaccgag
     2941 tttgggatca cctcagataa attgctgctg gcttggcggg agatggagca ggctgtggag
     3001 cagtgtgggc gggagaatga cgtgaagcat ctagtagagc ggatgatggc actgcagact
     3061 gacattgtgg acctgcagag gagcccgatg ggtcggaagc aggggggcac cctggatgac
     3121 ctagaggaac aagcgaggga gctctaccga agactcaggg agaagccaag agaccaaagg
     3181 acagaaggtg acagccagga gatggtacgg ctgctgcttc aggcaatcca aagctttgag
     3241 aagaaagttc gggtgattta tacacagctc agtaagaccg tggtttgtaa gcagaaggca
     3301 ctggagttgc tgcccaaggt agaagaggta gtgagcctta tgaacgagga cgagaggacc
     3361 gtggtccggc ttcaggagaa gcggcagaag gaactctgga acctcctgaa gatcgcctgt
     3421 agcaaagtcc gaggtcccgt gagtggaagc ccagacagca tgaatgtgtc tcgactcagt
     3481 caccctggtc agctaatgtc ccagccttcc agtgcctgtg acagcttacc tgaatcagac
     3541 aagaaaagtg aagaactggt ggccgaagcc cacgccctct gctcccggct agaaagtgcg
     3601 ctgcaggaca ctgtgaagga gcaagacaga agcttcacga ctctagactg gagctggtta
     3661 cagatggagg atgaagaaag gtgtagcctg gagcaggcct gtgacaatca ctagataact
     3721 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3781 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3841 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3901 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     3961 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4021 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4081 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4141 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4201 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4261 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4321 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4381 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     4441 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4501 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4561 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4621 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4681 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4741 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4801 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4861 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4921 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4981 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5041 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5101 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5161 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5221 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5281 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5341 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5401 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5461 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5521 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5581 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5641 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5701 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5761 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5821 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5881 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5941 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     6001 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     6061 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     6121 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     6181 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6241 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6301 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6361 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6421 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6481 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6961 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7021 ccgtattacc gccatgcat
//
