LOCUS       IKKalpha_(S176E,        9880 bp DNA     circular SYN 13-MAY-2021
DEFINITION  when used to produce lentivirus, express physiological levels of 
            insert .
ACCESSION   .
VERSION     .
KEYWORDS    IKKalpha (S176E,S180E)-pcw107-V5.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9880)
  AUTHORS   Martz CA, Ottina KA, Singleton KR, Jasper JS, Wardell SE, 
            Peraza-Penton A, Anderson GR, Winter PS, Wang T, Alley HM, Kwong LN,
            Cooper ZA, Tetzlaff M, Chen PL, Rathmell JC, Flaherty KT, Wargo JA, 
            McDonnell DP, Sabatini DM, Wood KC
  TITLE     Systematic identification of signaling pathways with potential to 
            confer anticancer drug resistance.
  JOURNAL   Sci Signal. 2014 Dec 23;7(357):ra121. doi: 
            10.1126/scisignal.aaa1877.
  PUBMED    25538079
REFERENCE   2  (bases 1 to 9880)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9880)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci
            Signal."; date: "2014-12-23"; pages: "
            10.1126/scisignal.aaa1877"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     promoter        1539..1749
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     CDS             1789..2385
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    2434..2551
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2606..3109
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     protein_bind    3130..3154
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3172..5406
                     /gene="CHUK"
                     /label=CHUK
                     /note="Inhibitor of nuclear factor kappa-B kinase subunit
                     alpha from Homo sapiens. Accession#: O15111"
     protein_bind    complement(5411..5431)
                     /label=attB2
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     CDS             5440..5481
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5506..6094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6166..6399
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6477..6611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6638..6773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6794..6812)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6822..6838)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6980..7435
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7461..7565
                     /label=AmpR promoter
     CDS             7566..8423
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8597..9185
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9339..9356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9473..9494
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9509..9539
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9547..9563
                     /label=lac operator
                     /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)."
     primer_bind     9571..9587
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9608..9626
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9654..9880
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccatgcattg catctcaatt
     1501 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1561 tgcgtctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1621 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1681 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1741 gcctaggctt ttgcaaaaag ctttctagag gtaccaccgg agcttaccat gaccgagtac
     1801 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     1861 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     1921 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     1981 tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag cgtcgaagcg
     2041 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     2101 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     2161 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     2221 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     2281 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     2341 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccagtta acgaattcta
     2401 gatcttgaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     2461 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     2521 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     2581 actttggcgc cggctcgagg cctgcaggtt gcgccttttc caaggcagcc ctgggtttgc
     2641 gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc
     2701 tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg
     2761 ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc
     2821 gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg
     2881 ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc
     2941 agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt
     3001 agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac
     3061 gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg ggctagcatc
     3121 gatggatcaa caagtttgta caaaaaagtt ggcccgggcg gcggcggcac catggagcgg
     3181 cccccggggc tgcggccggg cgcgggcggg ccctgggaga tgcgggagcg gctgggcacc
     3241 ggcggcttcg ggaacgtctg tctgtaccag catcgggaac ttgatctcaa aatagcaatt
     3301 aagtcttgtc gcctagagct aagtaccaaa aacagagaac gatggtgcca tgaaatccag
     3361 attatgaaga agttgaacca tgccaatgtt gtaaaggcct gtgatgttcc tgaagaattg
     3421 aatattttga ttcatgatgt gcctcttcta gcaatggaat actgttctgg aggagatctc
     3481 cgaaagctgc tcaacaaacc agaaaattgt tgtggactta aagaaagcca gatactttct
     3541 ttactaagtg atatagggtc tgggattcga tatttgcatg aaaacaaaat tatacatcga
     3601 gatctaaaac ctgaaaacat agttcttcag gatgttggtg gaaagataat acataaaata
     3661 attgatctgg gatatgccaa agatgttgat caaggagaac tgtgtacaga atttgtggga
     3721 acactgcagt atctggcccc agagctcttt gagaataagc cttacacagc cactgttgat
     3781 tattggagct ttgggaccat ggtatttgaa tgtattgctg gatataggcc ttttttgcat
     3841 catctgcagc catttacctg gcatgagaag attaagaaga aggatccaaa gtgtatattt
     3901 gcatgtgaag agatgtcagg agaagttcgg tttagtagcc atttacctca accaaatagc
     3961 ctttgtagtt taatagtaga acccatggaa aactggctac agttgatgtt gaattgggac
     4021 cctcagcaga gaggaggacc tgttgacctt actttgaagc agccaagatg ttttgtatta
     4081 atggatcaca ttttgaattt gaagatagta cacatcctaa atatgacttc tgcaaagata
     4141 atttcttttc tgttaccacc tgatgaaagt cttcattcat tacagtctcg tattgagcgt
     4201 gaaactggaa taaatactgg ttctcaagaa cttctttcag agacaggaat ttctctggat
     4261 cctcggaaac cagcctctca atgtgttcta gatggagtta gaggctgtga tagctatatg
     4321 gtttatttgt ttgataaaag taaaactgta tatgaagggc catttgcttc cagaagttta
     4381 tctgattgtg taaattatat tgtacaggac agcaaaatac agcttccaat tatacagctg
     4441 cgtaaagcgt gggctgaagc agtgcactat gtgtctggac taaaagaaga ctatagcagg
     4501 ctctttcagg gacaaagggc agcaatgtta agtcttctta gatataatgc taacttaaca
     4561 aaaatgaaga acactttgat ctcagcatca caacaactga aagctaaatt ggagtttttt
     4621 cacaaaagca ttcagcttga cttggagaga tacagcgagc agatgacgta tgggatatct
     4681 tcagaaaaaa tgctaaaagc atggaaagaa atggaagaaa aggccatcca ctatgctgag
     4741 gttggtgtca ttggatacct ggaggatcag attatgtctt tgcatgctga aatcatggag
     4801 ctacagaaga gcccctatgg aagacgtcag ggagacttga tggaatctct ggaacagcgt
     4861 gccattgatc tatataagca gttaaaacac agaccttcag atcactccta cagtgacagc
     4921 acagagatgg tgaaaatcat tgtgcacact gtgcagagtc aggaccgtgt gctcaaggag
     4981 ctgtttggtc atttgagcaa gttgttgggc tgtaagcaga agattattga tctactccct
     5041 aaggtggaag tggccctcag taatatcaaa gaagctgaca atactgtcat gttcatgcag
     5101 ggaaaaaggc agaaagaaat atggcatctc cttaaaattg cctgtacaca gagttctgcc
     5161 cggtcccttg taggatccag tctagaaggt gcagtaaccc ctcagacatc agcatggctg
     5221 cccccgactt cagcagaaca tgatcattct ctgtcatgtg tggtaactcc tcaagatggg
     5281 gagacttcag cacaaatgat agaagaaaat ttgaactgcc ttggccattt aagcactatt
     5341 attcatgagg caaatgagga acagggcaat agtatgatga atcttgattg gagttggtta
     5401 acagaattgc caactttctt gtacaaagtg gttaccggtg gtaagcctat ccctaaccct
     5461 ctcctcggtc tcgattctac gtagtaatga ccgcgttaag tcgacaatca acctctggat
     5521 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5581 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5641 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5701 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5761 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5821 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5881 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5941 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     6001 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6061 acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca
     6121 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     6181 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     6241 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     6301 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     6361 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc
     6421 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     6481 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6541 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6601 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6721 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac
     6781 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     6841 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     6901 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     6961 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7021 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     7081 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7141 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     7201 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     7261 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     7321 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     7381 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     7441 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7501 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7561 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7621 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7681 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7741 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7801 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7861 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7921 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7981 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     8041 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     8101 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     8161 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     8221 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     8281 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8341 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8401 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8461 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     8521 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8581 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8641 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8701 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8761 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8821 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8881 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8941 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     9001 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     9061 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     9121 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     9181 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     9241 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     9301 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     9361 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     9421 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     9481 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     9541 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     9601 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
     9661 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
     9721 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
     9781 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
     9841 cagagatatt gtatttaagt gcctagctcg atacataaac
//
