LOCUS       sequence-334992-        6274 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express IKK-epsilon v2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6274)
  TITLE     IKK-epsilon isoform switching governs the immune response against 
            EV71 infection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6274)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: F-IKK-epsilon v2
FEATURES             Location/Qualifiers
     source          1..6274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..319
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     276..296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(321..425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      452..907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(539..558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     749..770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    914..1035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(925..944)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     979..998
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1309..1328
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1309..1327
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1383..1399
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1384..1400
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3398..3421)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3421..3430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3465..3483)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(3529..3732)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3558..3578)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3733..4036
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4211..4799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4291..4310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5128..5175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5200..5219
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5407..6201)
                     /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             complement(5407..6201)
                     /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(5518..5537)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6128..6147
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6236..6274,1..319))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6250..6274,1..111)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
       61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      301 ctggttcttt ccgcctcagg attcttcctt tttcaatatt attgaagcat ttatcagggt
      361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      421 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
      481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      661 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      781 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      841 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      901 acaatttacg cgttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      961 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1021 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1081 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     1141 catgaacaga ctgtgaggac tgaggggcct gaaatgagcc ttgggactgt gaatctaaaa
     1201 tacacaaaca attagaatca gtagtttaac acattataca cttaaaaatt ggatctccat
     1261 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcga gtgaattgta atacgactca
     1321 ctatagggcg aattgggtac acttacctgg taccttaatt aattaaggta ccgggccccc
     1381 cctcgaggtc gacggtatcg ataagctttc agatgccagc agcttcatcc cctcgcagag
     1441 ctcttgcatg cttgctgaga ctctcctgga ccccctgggc ttctgtgtta caggcagcca
     1501 cagaacagcc aaccaggcgc aggtggttcc tggtctcgtg caccaccctc atcctcttgc
     1561 cgtgtgtgac taaggacgct tgatacttct gcacgcactc ctcctggaac acctgcagga
     1621 gtcttttggc taaatgactg aaattcacct tatccagctt gtgaatctgc tcctcgttgt
     1681 agccaagccc tggcctcatc ctagacttct tgaactgttt gtagatgaag ttcatcttgt
     1741 ccaaacagca ctgaatctgc tggatgcttc tgtcctcatg tacctgatcc cggctcttca
     1801 ccagctcccg gttcaggctg ctcaggctct cctgggtctc cgtgatattt tgggagcatc
     1861 tggagaggac ctccgctaga gtccgcagcc tggacctcag ttctgcagcc gccttcagtt
     1921 cctggatctc aggcgttcca gccacgctgc tgaacctctc agttcccagg ctgctgctga
     1981 ggtagaggag ggatgtcctt gccacttcca gagtccgtct gcatgtggcc tggagcacct
     2041 ccatgaccca gtgcagcccc cgaaacatta gctcctgccc atccagcagg gcccgtgcca
     2101 gccgcagggc ctggtagccg gcgcccaaca cgcccttggc agtgttgtaa tccgcctgca
     2161 ggtccacttt ggggacgaac ttggggacgt ccagagcagg gtccctgaag gccagcccct
     2221 tagggatggc tgtgctgaag agggtcaggg ggctgcttgc cgtcgtgtgg gcgatgtgct
     2281 gtgctgagac gctgggctcg aggacacaga ggtgaccctc aaagaggtac tcctggtgtc
     2341 ggggggccac actggtctgc ttgtgcacgg cctcctggaa aatggctatc gtgttgtggg
     2401 catggatata gatgtggtgc aggactgcct gggacaggga gaagacatgg acgacaactc
     2461 gctgcaggat gtcactggtc tccgcaaaga actggccgaa gccccagcac ttggcctgct
     2521 ccacctccag gatgttggcc aggatgggca ccagctggct ctgcagcccc agtgacagct
     2581 ggcaggtgat ggggagggtg tagctccact ccaggggccc gttctcccgc ctctgggcac
     2641 ctgcaatggc cccagccggc ttctccgtgg tgatccggta catgatctcc ttgttccgcc
     2701 gtggcccacc aaaggggatg aagggcaggc tgccagtggc tgcatggtac aaggtcactc
     2761 caatgctcca gagatccaca gtcaccccga acgctttttg ctggggcttt cgaagcaccg
     2821 cccgctcata catgtcggga tgcaggtact cctcagtccc atagaccgag acgaacttct
     2881 catcatcatc cagctcccgg gcagcgccga agtctgtcag cttgtagatg ctctgcccct
     2941 cctcccctac gaggcgcatg atgttccccg gcttgatgtc gcgatgcaca atgccgttct
     3001 cccgcaggtg gttcatgctg gccaccacac agcgcagcac caccaggaac tcatcctcag
     3061 gcagcccaaa ggcattctca gggctctcca gcacactcag caggctccca ctggagcagt
     3121 actccatcac cagtaccttc tgccggcttc cgcccgtctc ctccaccgca aagagcttga
     3181 cgatgttctg gtggttcagc ttccgcagga cctcaaactc cctcacctgc acctcgcggg
     3241 gccgcaggta gctggtagtg ttgaagacct tcacagcaac cagctctccg gatttcttgt
     3301 tgcgggcctt gtacacactg gcagtggccc cctgccccag caggtcatct gtgtgccaca
     3361 ggtaattggc tgtgctctgc atggatccgc ccgggctctt atcgtcgtca tccttgtaat
     3421 ccatggtggc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     3481 atttcgagct tggcgtaatc gctagcggat ctgacggttc actaaaccag ctctgcttat
     3541 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     3601 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     3661 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     3721 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     3781 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     3841 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     3901 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     3961 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     4021 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     4081 ttactattaa taactaatgc atggcggtaa tacggttatc cacagaatca ggggataacg
     4141 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4201 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4261 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4321 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4381 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4441 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4501 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4561 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4621 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4681 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4741 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4801 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4861 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4921 gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg
     4981 ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa
     5041 aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc
     5101 agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct
     5161 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     5221 ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     5281 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     5341 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     5401 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     5461 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagtt cttcagcaat
     5521 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     5581 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     5641 ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     5701 tggcgcgagc ccctgatgtt cttcgtccag atcatcctga tcgacaagac cggcttccat
     5761 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     5821 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     5881 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     5941 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     6001 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     6061 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     6121 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     6181 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc
     6241 aaaagcctag gcctccaaaa aagcctcctc acta
//
