Opie2-PuroR vector (V012836)

Basic Vector Information

Expresses PuroR and dsRed.

      • Vector Name:
      • Opie2-PuroR
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 8646 bp
      • Type:
      • Insect Cell Vectors
      • Source/Author:
      • Omar Akbari
      • Promoter:
      • Opie2
      • Fusion Tag:
      • DsRed1
      • Growth Strain(s):
      • DH5alpha
      • Growth Temperature:
      • 37℃

Opie2-PuroR vector Vector Map

Opie2-PuroR8646 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400L4440CAP binding sitelac promoterlac operatorM13 revpiggyBac left (5') inverted repeatSV40 poly(A) signalDsRed1IE1 promoterhr5 enhancerPuroRSV40 poly(A) signalpiggyBac right (3') inverted repeatM13 ForwardpRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRori

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

Opie2-PuroR vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                8646 bp DNA     circular SYN 15-JAN-2024
DEFINITION  Expresses PuroR and dsRed..
ACCESSION   .
VERSION     .
KEYWORDS    Opie2-PuroR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8646)
  AUTHORS   Kandul NP, Liu J, Hsu AD, Hay BA, Akbari OS
  TITLE     A drug-inducible sex-separation technique for insects.
  JOURNAL   Nat Commun. 2020 Apr 30;11(1):2106. doi: 10.1038/s41467-020-16020-2.
  PUBMED    32355156
REFERENCE   2  (bases 1 to 8646)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-04-30"; volume: "11(1):2106. doi"; pages: " 
            10.1038/s41467-020-16020-2"
FEATURES             Location/Qualifiers
     source          1..8646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..77
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    194..215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        230..260
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    268..284
                     /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     273..295
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     292..308
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     292..308
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   489..523
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal
                     repeat sequence (ITR)"
     polyA_signal    1212..1333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1454..2134)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1516..1539)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1714..1733)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1931..1951
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     promoter        complement(2145..2736)
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus
                     Autographa californica"
     enhancer        2740..3222
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     CDS             3785..4384
                     /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(3785..3804)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also
                     called puro-variant-R"
     primer_bind     4281..4301
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4402..4536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   5572..5634
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal
                     repeat sequence (ITR)"
     primer_bind     complement(6338..6355)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     complement(6338..6354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6347..6369)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6563..6582)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     6682..6704
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6742..6760)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        6828..6932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6933..7793
                     /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(7151..7170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7964..8552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8453..8472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"

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