LOCUS       sequence-241040-        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 flag
FEATURES             Location/Qualifiers
     source          1..7550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..486
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        487..690
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     641..661
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     735..754
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        735..753
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             776..799
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2975..2993)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2976..2993)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3013..3030)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3019..3243
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3289..3717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3376..3395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3586..3607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3726..3746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3731..4060
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3911..4046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3973..3992
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4127..4921
                     /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(4181..4200)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4791..4810
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5095..5216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5132..5151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5186..5205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5265..5281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5265..5281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5278..5300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5289..5305
                     /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(5313..5319,5320..5337,5338..5343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5358..5379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5496..5513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5667..6255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5747..5766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6426..7217,7218..7286))
                     /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     7049..7068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7287..7391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7466..7485)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
       61 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      121 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      181 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      241 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      301 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      361 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      421 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      481 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      541 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      601 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      661 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      721 tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta ccatggacta
      781 caaagacgat gacgataagg atccggagat gcagagcaca gccaattacc tgtggcacac
      841 agatgacctg ctggggcagg gggccactgc cagtgtgtac aaggcccgca acaagaaatc
      901 cggagagctg gttgctgtga aggtcttcaa cactaccagc tacctgcggc cccgcgaggt
      961 gcaggtgagg gagtttgagg tcctgcggaa gctgaaccac cagaacatcg tcaagctctt
     1021 tgcggtggag gagacgggcg gaagccggca gaaggtactg gtgatggagt actgctccag
     1081 tgggagcctg ctgagtgtgc tggagagccc tgagaatgcc tttgggctgc ctgaggatga
     1141 gttcctggtg gtgctgcgct gtgtggtggc cggcatgaac cacctgcggg agaacggcat
     1201 tgtgcatcgc gacatcaagc cggggaacat catgcgcctc gtaggggagg aggggcagag
     1261 catctacaag ctgacagact tcggcgctgc ccgggagctg gatgatgatg agaagttcgt
     1321 ctcggtctat gggactgagg agtacctgca tcccgacatg tatgagcggg cggtgcttcg
     1381 aaagccccag cagaaagcgt tcggggtgac tgtggatctc tggagcattg gagtgacctt
     1441 gtaccatgca gccactggca gcctgccctt catccccttt ggtgggccac ggcggaacaa
     1501 ggagatcatg taccggatca ccacggagaa gccggctggg gccattgcag gtgcccagag
     1561 gcgggagaac gggcccctgg agtggagcta caccctcccc atcacctgcc agctgtcact
     1621 ggggctgcag agccagctgg tgcccatcct ggccaacatc ctggaggtgg agcaggccaa
     1681 gtgctggggc ttcgaccagt tctttgcgga gaccagtgac atcctgcagc gagttgtcgt
     1741 ccatgtcttc tccctgtccc aggcagtcct gcaccacatc tatatccatg cccacaacac
     1801 gatagccatt ttccaggagg ccgtgcacaa gcagaccagt gtggcccccc gacaccagga
     1861 gtacctcttt gagggtcacc tctgtgtcct cgagcccagc gtctcagcac agcacatcgc
     1921 ccacacgacg gcaagcagcc ccctgaccct cttcagcaca gccatcccta aggggctggc
     1981 cttcagggac cctgctctgg acgtccccaa gttcgtcccc aaagtggacc tgcaggcgga
     2041 ttacaacact gccaagggcg tgttgggcgc cggctaccag gccctgcggc tggcacgggc
     2101 cctgctggat gggcaggagc taatgtttcg ggggctgcac tgggtcatgg aggtgctcca
     2161 ggccacatgc agacggactc tggaagtggc aaggacatcc ctcctctacc tcagcagcag
     2221 cctgggaact gagaggttca gcagcgtggc tggaacgcct gagatccagg aactgaaggc
     2281 ggctgcagaa ctgaggtcca ggctgcggac tctagcggag gtcctctcca gatgctccca
     2341 aaatatcacg gagacccagg agagcctgag cagcctgaac cgggagctgg tgaagagccg
     2401 ggatcaggta catgaggaca gaagcatcca gcagattcag tgctgtttgg acaagatgaa
     2461 cttcatctac aaacagttca agaagtctag gatgaggcca gggcttggct acaacgagga
     2521 gcagattcac aagctggata aggtgaattt cagtcattta gccaaaagac tcctgcaggt
     2581 gttccaggag gagtgcgtgc agaagtatca agcgtcctta gtcacacacg gcaagaggat
     2641 gagggtggtg cacgagacca ggaaccacct gcgcctggtt ggctgttctg tggctgcctg
     2701 taacacagaa gcccaggggg tccaggagag tctcagcaag ctcctggaag agctatctca
     2761 ccagctcctt caggaccgag caaagggggc tcaggcctcg ccgcctccca tagctcctta
     2821 ccccagccct acacgaaagg acctgcttct ccacatgcaa gagctctgcg aggggatgaa
     2881 gctgctggca tctgacctcc tggacaacaa ccgcatcatc gaacggctaa atagagtccc
     2941 agcacctcct gatgtctgat ctagagggcc ctattctata gtgtcaccta aatgctagag
     3001 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3061 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3121 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3181 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3241 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3301 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3361 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3421 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3481 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3541 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3601 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3661 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3721 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3781 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3841 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3901 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3961 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     4021 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     4081 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4141 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4201 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4261 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4321 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4381 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4441 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4501 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4561 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4621 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4681 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4741 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4801 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4861 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4921 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4981 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5041 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5101 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5161 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5221 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5281 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5341 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5401 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5461 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6541 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7381 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     7441 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     7501 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
//
