LOCUS Exported 7083 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLKO.1.sh.beta-catenin.1248
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7083)
AUTHORS Firestein R, Bass AJ, Kim SY, Dunn IF, Silver SJ, Guney I, Freed E,
Ligon AH, Vena N, Ogino S, Chheda MG, Tamayo P, Finn S, Shrestha Y,
Boehm JS, Jain S, Bojarski E, Mermel C, Barretina J, Chan JA,
Baselga J, Tabernero J, Root DE, Fuchs CS, Loda M, Shivdasani RA,
Meyerson M, Hahn WC
TITLE CDK8 is a colorectal cancer oncogene that regulates beta-catenin
activity.
JOURNAL Nature. 2008 Sep 25. 455(7212):547-51.
PUBMED 18794900
REFERENCE 2 (bases 1 to 7083)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
2008 Sep 25. 455(7212):547-51."
FEATURES Location/Qualifiers
source 1..7083
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 50..67
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 184..205
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 220..250
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 258..274
/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 263..285
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 282..298
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 282..298
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 317..337
/label=T3
/note="T3 promoter, forward primer"
promoter 319..337
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 365..591
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 592..772
/label=5 LTR (truncated)
/note="truncated 5 long terminal repeat (LTR) from HIV-1"
misc_feature 819..944
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1437..1670
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1855..1899
/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 2197..2437
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 2197..2217
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2368..2387
/label=LKO.1 5
/note="Human U6 promoter, forward primer"
misc_feature 2541..2658
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2707..3217
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 3136..3154
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 3239..3838
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3239..3258)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3735..3755
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3966..4199
/label=3 LTR (Delta-U3)
/note="self-inactivating 3 long terminal repeat (LTR) from
HIV-1"
polyA_signal 4271..4392
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4308..4327)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4362..4381
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4432..4567
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4494..4513
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4587..4606)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4588..4606)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4616..4633)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4616..4632)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4625..4647)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4774..5229
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4861..4880)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5071..5092
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5255..5359
/gene="bla"
/label=AmpR promoter
CDS 5360..6220
/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(5578..5597)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6391..6979
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6880..6899
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
