LOCUS       Exported                9880 bp ds-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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Signal."; date: "2014-12-23"; volume: "7(357):ra121. doi"; pages: " 
            10.1126/scisignal.aaa1877"
FEATURES             Location/Qualifiers
     source          1..9880
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        1539..1749
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        1561..1757
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1608..1743
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1789..2391
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(1789..1808)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2285..2305
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2434..2551
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2609..3109
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3028..3046
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     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"
     protein_bind    complement(5408..5432)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="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"
     primer_bind     complement(5559..5579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5977..5988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6166..6399
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6477..6598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6514..6533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6568..6587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6638..6773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6793..6812)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6794..6812)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6822..6839)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6822..6838)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6831..6853)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6980..7435
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7067..7086)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7277..7298
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7461..7565
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7566..8426
                     /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     complement(7784..7803)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8597..9185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9086..9105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9339..9356
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9473..9494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /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)."
     primer_bind     9552..9574
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9571..9587
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9571..9587
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9606..9626
                     /label=T3
                     /note="T3 promoter, forward primer"
     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"