pAPi vector (V015884)

LOCUS       62056_3160        7827 bp DNA     circular SYN 25-MAY-2020
DEFINITION  UNVERIFIED: Cloning vector pAPi, complete sequence.
ACCESSION   MK975251
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  AUTHORS   Foley SJ, Orr RG, Galotto G, Liu B, Vidali L.
  TITLE     Robust survival-based RNAi in plants using in tandem silencing of 
            adenine phosphoribosyltransferase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7827)
  AUTHORS   Foley SJ, Orr RG, Galotto G, Liu B, Vidali L.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAY-2019) Biology and Biotechnology, Worcester 
            Polytechnic Institute, 100 Institute Rd., Worcester, MA 01609, USA
REFERENCE   3  (bases 1 to 7827)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-MAY-2019) Biology and Biotechnology, Worcester Polytechnic 
            Institute, 100 Institute Rd., Worcester, MA 01609, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            GenBank staff is unable to verify sequence and/or annotation 
            provided by the submitter.
FEATURES             Location/Qualifiers
     source          1..7827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3195..3447
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3480..3824
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3854..4876
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4948..5124
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5681..5697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5705..5721)
                     /label=lac operator
                     /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)."
     promoter        complement(5729..5759)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5774..5795)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6083..6671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6845..7702)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7703..7807)
                     /label=AmpR promoter
     promoter        7827
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"