LOCUS Exported 13870 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS aMHC-eGFP-Rex-Neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13870)
AUTHORS Kita-Matsuo H, Barcova M, Prigozhina N, Salomonis N, Wei K, Jacot
JG, Nelson B, Spiering S, Haverslag R, Kim C, Talantova M, Bajpai R,
Calzolari D, Terskikh A, McCulloch AD, Price JH, Conklin BR, Chen
HS, Mercola M
TITLE Lentiviral vectors and protocols for creation of stable hESC lines
for fluorescent tracking and drug resistance selection of
cardiomyocytes.
JOURNAL PLoS ONE. 2009 . 4(4):e5046.
PUBMED 19352491
REFERENCE 2 (bases 1 to 13870)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE.
2009 . 4(4):e5046."
FEATURES Location/Qualifiers
source 1..13870
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 27..131
/gene="bla"
/label=AmpR promoter
CDS 132..992
/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(350..369)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1163..1751
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1652..1671
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1905..1922
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2039..2060
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2075..2105
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2113..2129
/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 2118..2140
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2137..2153
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2137..2153
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 2172..2192
/label=T3
/note="T3 promoter, forward primer"
promoter 2174..2192
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
LTR 2448..2628
/label=5 LTR (truncated)
/note="truncated 5 long terminal repeat (LTR) from HIV-1"
misc_feature 2672..2797
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3290..3523
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 3708..3752
/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"
misc_feature 4019..4135
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 4181..4197
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4181..4197
/label=pBluescriptSK
/note="For pBluescript vector"
regulatory 9664..9673
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 9670..10389
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 9670..10386
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(9715..9736)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(9976..9995)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 10323..10344
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 10405..10993
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(10458..10478)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(10876..10887)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
regulatory 11729..11738
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 11735..12538
/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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(11798..11817)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 12408..12427
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
LTR 12611..12844
/label=3 LTR (Delta-U3)
/note="self-inactivating 3 long terminal repeat (LTR) from
HIV-1"
polyA_signal 12916..13037
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(12953..12972)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 13007..13026
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 13056..13191
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 13118..13137
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(13223..13242)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(13224..13242)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(13252..13269)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(13252..13268)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(13261..13283)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 13413..13868
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(13500..13519)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 13710..13731
/label=F1ori-F
/note="F1 origin, forward primer"
