7TFP CDH1 reporter vector (V006706)

Basic Vector Information

      • Vector Name:
      • 7TFP CDH1 reporter
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9174 bp
      • Type:
      • Mammalian Expression, Lentiviral, Luciferase
      • Selection Marker:
      • Puromycin
      • Copy Number:
      • High Copy
      • Promoter:
      • CDH1
      • Cloning Method:
      • Restriction Enzyme
      • 3' Primer:
      • LucNRev

7TFP CDH1 reporter vector Vector Map

7TFP CDH1 reporter9174 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800minPKozak sequencepBABE 3'PuroRWPRE3' LTR (Delta-U3)SV40 poly(A) signalSV40 oriT7M13 Forwardf1 oriAmpR promoterAmpRoriL4440CAP binding sitelac promoterlac operatorM13 revT3RSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptidecPPT/CTS

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 ℃.

7TFP CDH1 reporter vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9174 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  lentiviral luciferase reporter containing CDH1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    7TFP CDH1 reporter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9174)
  AUTHORS   Pattabiraman DR, Bierie B, Kober KI, Thiru P, Krall JA, Zill C, 
            Reinhardt F, Tam WL, Weinberg RA
  TITLE     Activation of PKA leads to mesenchymal-to-epithelial transition and 
            loss of tumor-initiating ability.
  JOURNAL   Science. 2016 Mar 4;351(6277):aad3680. doi: 10.1126/science.aad3680.
  PUBMED    26941323
REFERENCE   2  (bases 1 to 9174)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2016-03-4"; volume: "351(6277):aad3680. doi"; pages: " 
            10.1126/science.aad3680"
FEATURES             Location/Qualifiers
     source          1..9174
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        325..356
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      412..421
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             418..2070
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(484..502)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2089..2109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2094..2423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2274..2409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2433..3032
                     /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(2433..2452)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2929..2949
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3099..3687
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3152..3172)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3570..3581)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3774..4007
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4079..4200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4116..4135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4170..4189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4240..4375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4395..4414)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4396..4414)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4424..4441)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4424..4440)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4433..4455)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4582..5037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4669..4688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4879..4900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5063..5167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5168..6028
                     /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(5386..5405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6199..6787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6688..6707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6941..6958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7075..7096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7111..7141
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7149..7165
                     /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     7154..7176
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7173..7189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7173..7189
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7208..7228
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7210..7228
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7256..7482
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7483..7663
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7710..7835
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8328..8561
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8746..8790
                     /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    9057..9174
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"

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