LOCUS       sequence-366329-        7550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7550)
  AUTHORS   Fitzgerald KA, McWhirter SM, Faia KL, Rowe DC, Latz E, Golenbock DT,
            Coyle AJ, Liao SM, Maniatis T
  TITLE     IKKepsilon and TBK1 are essential components of the IRF3 signaling 
            pathway.
  JOURNAL   Nat Immunol. 2003 May;4(5):491-6.
  PUBMED    12692549
REFERENCE   2  (bases 1 to 7550)
  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 
            Immunol."; date: "2003-05"; volume: "4(5)"; pages: "491-6"
COMMENT     Sequence Label: pcDNA3 IKKE K38A flag
FEATURES             Location/Qualifiers
     source          1..7550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..55)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        40..369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      220..355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     282..301
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             436..1230
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(490..509)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1100..1119
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1404..1525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1441..1460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1495..1514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1574..1590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1574..1590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1587..1609)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1598..1614
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1622..1628,1629..1646,1647..1652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1667..1688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1805..1822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1976..2564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2056..2075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2735..3526,3527..3595))
                     /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     3358..3377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3596..3700)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3775..3794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3966..4345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4346..4549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4500..4520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4594..4613
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4594..4612
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4635..4658
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(6834..6852)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6835..6852)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6872..6889)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6878..7102
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7148..7550,1..26)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7235..7254)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7445..7466
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
       61 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
      121 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      181 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
      241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
      301 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
      361 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
      421 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
      481 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
      541 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
      601 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
      661 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
      721 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
      781 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
      841 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
      901 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
      961 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     1021 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     1081 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     1141 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     1201 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     1261 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     1321 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     1381 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     1441 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     1501 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     1561 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1621 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1681 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1741 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1801 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1861 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1921 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1981 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2041 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2101 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2161 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2221 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2281 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2341 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2401 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2461 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2521 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2581 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2641 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2701 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2761 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2821 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2881 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2941 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3001 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3061 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3121 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3181 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3241 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3301 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3361 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3421 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3481 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3541 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3601 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3661 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3721 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     3781 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     3841 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     3901 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     3961 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     4021 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     4081 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     4141 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     4201 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     4261 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     4321 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     4381 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     4441 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     4501 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
     4561 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     4621 agcttggtac catggactac aaagacgatg acgataagga tccggagatg cagagcacag
     4681 ccaattacct gtggcacaca gatgacctgc tggggcaggg ggccactgcc agtgtgtaca
     4741 aggcccgcaa caagaaatcc ggagagctgg ttgctgtggc ggtcttcaac actaccagct
     4801 acctgcggcc ccgcgaggtg caggtgaggg agtttgaggt cctgcggaag ctgaaccacc
     4861 agaacatcgt caagctcttt gcggtggagg agacgggcgg aagccggcag aaggtactgg
     4921 tgatggagta ctgctccagt gggagcctgc tgagtgtgct ggagagccct gagaatgcct
     4981 ttgggctgcc tgaggatgag ttcctggtgg tgctgcgctg tgtggtggcc ggcatgaacc
     5041 acctgcggga gaacggcatt gtgcatcgcg acatcaagcc ggggaacatc atgcgcctcg
     5101 taggggagga ggggcagagc atctacaagc tgacagactt cggcgctgcc cgggagctgg
     5161 atgatgatga gaagttcgtc tcggtctatg ggactgagga gtacctgcat cccgacatgt
     5221 atgagcgggc ggtgcttcga aagccccagc agaaagcgtt cggggtgact gtggatctct
     5281 ggagcattgg agtgaccttg taccatgcag ccactggcag cctgcccttc atcccctttg
     5341 gtgggccacg gcggaacaag gagatcatgt accggatcac cacggagaag ccggctgggg
     5401 ccattgcagg tgcccagagg cgggagaacg ggcccctgga gtggagctac accctcccca
     5461 tcacctgcca gctgtcactg gggctgcaga gccagctggt gcccatcctg gccaacatcc
     5521 tggaggtgga gcaggccaag tgctggggct tcgaccagtt ctttgcggag accagtgaca
     5581 tcctgcagcg agttgtcgtc catgtcttct ccctgtccca ggcagtcctg caccacatct
     5641 atatccatgc ccacaacacg atagccattt tccaggaggc cgtgcacaag cagaccagtg
     5701 tggccccccg acaccaggag tacctctttg agggtcacct ctgtgtcctc gagcccagcg
     5761 tctcagcaca gcacatcgcc cacacgacgg caagcagccc cctgaccctc ttcagcacag
     5821 ccatccctaa ggggctggcc ttcagggacc ctgctctgga cgtccccaag ttcgtcccca
     5881 aagtggacct gcaggcggat tacaacactg ccaagggcgt gttgggcgcc ggctaccagg
     5941 ccctgcggct ggcacgggcc ctgctggatg ggcaggagct aatgtttcgg gggctgcact
     6001 gggtcatgga ggtgctccag gccacatgca gacggactct ggaagtggca aggacatccc
     6061 tcctctacct cagcagcagc ctgggaactg agaggttcag cagcgtggct ggaacgcctg
     6121 agatccagga actgaaggcg gctgcagaac tgaggtccag gctgcggact ctagcggagg
     6181 tcctctccag atgctcccaa aatatcacgg agacccagga gagcctgagc agcctgaacc
     6241 gggagctggt gaagagccgg gatcaggtac atgaggacag aagcatccag cagattcagt
     6301 gctgtttgga caagatgaac ttcatctaca aacagttcaa gaagtctagg atgaggccag
     6361 ggcttggcta caacgaggag cagattcaca agctggataa ggtgaatttc agtcatttag
     6421 ccaaaagact cctgcaggtg ttccaggagg agtgcgtgca gaagtatcaa gcgtccttag
     6481 tcacacacgg caagaggatg agggtggtgc acgagaccag gaaccacctg cgcctggttg
     6541 gctgttctgt ggctgcctgt aacacagaag cccagggggt ccaggagagt ctcagcaagc
     6601 tcctggaaga gctatctcac cagctccttc aggaccgagc aaagggggct caggcctcgc
     6661 cgcctcccat agctccttac cccagcccta cacgaaagga cctgcttctc cacatgcaag
     6721 agctctgcga ggggatgaag ctgctggcat ctgacctcct ggacaacaac cgcatcatcg
     6781 aacggctaaa tagagtccca gcacctcctg atgtctgatc tagagggccc tattctatag
     6841 tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     6901 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     6961 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     7021 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     7081 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     7141 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     7201 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     7261 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     7321 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     7381 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     7441 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     7501 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
//
