Basic Vector Information
Plasmid for expressing evolved and engineered Bxb1 in the mammalian cells
- Vector Name:
- eeBxb1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5801 bp
- Type:
- Mammalian Expression Vectors
- Promoter:
- CMV
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
eeBxb1 vector Map
eeBxb1 vector Sequence
LOCUS Exported 5801 bp DNA circular SYN 06-SEP-2024 DEFINITION Plasmid for expressing evolved and engineered Bxb1 in the mammalian cells. ACCESSION . VERSION . KEYWORDS eeBxb1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5801) AUTHORS Pandey S, Gao XD, Krasnow NA, McElroy A, Tao YA, Duby JE, Steinbeck BJ, McCreary J, Pierce SE, Tolar J, Meissner TB, Chaikof EL, Osborn MJ, Liu DR TITLE Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases and prime editing. JOURNAL Nat Biomed Eng. 2024 Jun 10. doi: 10.1038/s41551-024-01227-1. PUBMED 38858586 REFERENCE 2 (bases 1 to 5801) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Biomed Eng. 2024 Jun 10. doi: 10.1038/s41551-024-01227-1." FEATURES Location/Qualifiers source 1..5801 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 743..760 /label=L4440 /note="L4440 vector, forward primer" promoter complement(834..938) /gene="bla" /label=AmpR promoter primer_bind 1006..1024 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1051..1081 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 1089..1105 /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 1094..1116 /label=M13/pUC Reverse /note="In lacZ gene" enhancer 1135..1438 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1439..1642 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1593..1613 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 1639..1663 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1758..1776 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1758..1775 /label=SP6 /note="SP6 promoter, forward primer" CDS 1814..3316 /codon_start=1 /transl_table=11 /gene="Bxb1" /product="Bxb1 integrase" /label=Bxb1 integrase /note="Bxb1 large serine integrase from Mycobacterium bacteriophage Bxb1 (Ghosh et al. 2003)." /protein_id="AFF61396.1" /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD VSGAVDPFDRKRRPNLARWLAFEEQPFDAIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG REPQGRKWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAIELAEVNAELVDLTSLIGSPAYRAG SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS" polyA_signal 3728..3862 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3765..3784) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3819..3838 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3988..4442 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4075..4094) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4284..4305 /label=F1ori-F /note="F1 origin, forward primer" CDS 4729..5589 /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(4947..4966) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" protein_bind complement(5691..5724) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)."
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