pTKIP-neo vector (V002618)

LOCUS       40924_43553        4290 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Donor vector pTKIP-neo, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4290)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Site-specific chromosomal integration of large synthetic constructs
  JOURNAL   Nucleic Acids Res. 38 (6), E92 (2010)
  PUBMED    20047970
REFERENCE   2  (bases 1 to 4290)
  AUTHORS   Kuhlman TE, Cox EC.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-DEC-2009) Molecular Biology, Princeton University, 
            Washington Rd., Princeton, NJ 08544, USA
REFERENCE   3  (bases 1 to 4290)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4290)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2010"; volume: "38"; issue: "6"; pages: "E92"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-DEC-2009) Molecular Biology, Princeton University, Washington 
            Rd., Princeton, NJ 08544, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             238..426
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    531..671
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(857..1445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1619..2476)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2477..2581)
                     /label=AmpR promoter
     misc_feature    2607..2624
                     /label=I-SceI recognition site
                     /note="I-SceI recognition site"
     misc_feature    2625..2649
                     /label=Landing Pad Region 1
                     /note="Landing Pad Region 1"
     primer_bind     2668..2684
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2706..2739
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        2803..3302
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        3314..3361
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3380..4180
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4214..4247
                     /label=FRT
                     /note="FRT"
     protein_bind    4214..4247
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     misc_feature    4248..4272
                     /label=Landing Pad Region 2
                     /note="Landing Pad Region 2"
     misc_feature    4273..4290
                     /label=I-SceI recognition site
                     /note="I-SceI recognition site"