LOCUS       sequence-140321-        7031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse eGFP-IKK-alpha in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7031)
  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 7031)
  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-alpha
FEATURES             Location/Qualifiers
     source          1..7031
                     /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"
     RBS             3353..3361
                     /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      3652..3661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3656..3665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3819..3940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3856..3875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3910..3929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3947..4402)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4084..4105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4296..4315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4429..4533
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4535..4892
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4535..4726
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4558..4578)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4743..4878
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4927..5721
                     /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             4927..5721
                     /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(4981..5000)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5591..5610
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5909..5928)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5953..6000
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6329..6917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6818..6837
                     /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 cgacctgcag
     1381 gcggccgcac tagtgattat ggagcggccc ccggggctgc ggccgggcgc gggcggcccc
     1441 tgggagatgc gggaacggct tggcaccggc ggtttcggga acgtcagtct gtaccagcac
     1501 cgggaacttg atctcaaaat agcaattaag tcttgtcgtt tagagctaag ttccaaaaac
     1561 agagagcgat ggtgccatga aatccagatc atgaaaaagt tggaccatgc gaatgttgta
     1621 aaggcctgtg atgtccctga ggaattgaac tttttaatta acgatgtgcc tcttctggca
     1681 atggagtact gttctggagg ggacctccgg aagctactca acaaaccaga aaattgttgt
     1741 ggacttaaag aaagccagat actttcttta ctgagtgaca taggatctgg gatccgatat
     1801 ctgcatgaaa acaaaattat acatcgagat ctaaaacctg aaaatatagt tcttcaagat
     1861 gttggtggga agacaataca taaaataatt gatttgggtt atgccaaaga tgttgatcaa
     1921 ggaagtctct gtacatcttt tgtgggaaca ttgcagtatt tggccccaga gctctttgaa
     1981 aataagccgt acacagccac tgtggattat tggagctttg ggaccatggt gtttgaatgt
     2041 attgctggat ataggccttt tttgcatcat ctgcagccat ttacctggca tgagaagatt
     2101 aagaagaaag atccaaagtg tatatttgca tgtgaagaga tgactggaga agttcggttt
     2161 agtagccatt tacctcagcc aaacagcctt tgtagtttaa tagtagagcc aatggaaagc
     2221 tggctccaat tgatgctgaa ttgggaccca cagcagagag ggggacctat tgatcttact
     2281 ttgaagcagc caagatgttt tgcattaatg gatcacattc tcaatttaaa gatagtgcac
     2341 atcctaaata tgacttctgc aaaaatcatt tcttttctgt taccatgtga tgaaagtctt
     2401 cattcactac agtctcgaat tgagcgtgaa acaggaataa atacaggttc tcaggagctt
     2461 ctgtcagaga cagggatttc tctggatcct cggaaaccag cctctcagtg tgttctagat
     2521 ggagttagag gctgtgatag ctacatggtt tatttgtttg ataaaagtaa gactgtatat
     2581 gaaggaccat ttgcatccag aagtttatct gattgtgtaa attatattgt acaagacagc
     2641 aaaatacaac tgccaattat acagctgcgg aaagtatggg ctgaagcagt gcactacgta
     2701 tctgggctaa aggaagacta cagcaggctc ttccagggac aaagagcagc aatgttaagt
     2761 cttcttagat ataatgctaa cttgacaaaa atgaagaata ctttgatctc agcatcacag
     2821 caactcaaag ctaaattgga gttttttcga aaaagcattc agcttgactt ggagagatat
     2881 agtgagcaga tgacttatgg gatatcttca gaaaaaatgt taaaagcatg gaaagaaatg
     2941 gaagaaaagg ccattcacta ttctgaggtt ggtgtcattg gttatcttga ggatcaaatt
     3001 atgtctttgc acactgaaat catggagctg cagaagagcc cctacggacg acgccaggga
     3061 gacttgatgg agtctctgga gcagcgtgcc attgatctct ataagcagct aaagcacaga
     3121 cctcctgatc acttgtacag cgacagcaca gagatggtga agatcatcgt gcacaccgtg
     3181 cagagtcagg accgtgttct caaggagctg tttggtcacc tgagcaagtt gttgggctgc
     3241 aagcagaaga ttattgatct actccccaag gtggaagtgg ccctcagtaa catcaaagaa
     3301 gctgacaata ctgtcatgtt tatgcaggga aagaggcaga aagaaatttg gcacctcctt
     3361 aaaattgcct gtacacagag ttctgcccgc tctcttgtag gatccagtct agaaggcaca
     3421 gtaacccctc cagtatcagc atggctgccc cctacattag cagaccgtga acatcctctg
     3481 acatatgtgg taactcctca agatggagag acgttagcac aaatgataga agaaaatctg
     3541 aactgtcttg gccatttaag tactattatt cgtgaagcaa atgaggacca gagcagtagt
     3601 ttgatgagtc ttgattggag ttggttagca gaatgaaatc gaattcccgc ggccgccatg
     3661 gcggccggga gcatgcgacg tcgggcccgg gatccaccgg atctagataa ctgatcataa
     3721 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3781 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3841 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3901 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3961 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4021 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4081 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4141 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4201 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4261 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4321 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4381 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4441 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4501 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4561 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4621 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4681 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4741 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4801 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4861 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4921 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4981 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5041 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5101 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5161 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5221 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5281 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5341 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5401 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5461 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5521 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5581 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5641 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5701 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5761 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5821 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5881 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5941 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6001 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6061 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6121 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     6181 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     6481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7021 ccgccatgca t
//
