Alpha-3000-CRgli2PSM vector (V010124)

Basic Vector Information

      • Vector Name:
      • Alpha-3000-CRgli2PSM
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 11126 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.

Alpha-3000-CRgli2PSM vector Vector Map

Alpha-3000-CRgli2PSM11126 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000Gli2SV40 poly(A) signalT7 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterM13 fwd

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 ℃.

Alpha-3000-CRgli2PSM vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               11126 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector Alpha-3000-CRgli2PSM, complete sequence.
ACCESSION   MF444684
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11126)
  AUTHORS   Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.
  TITLE     Gli2 is required for the induction and migration of Xenopus laevis 
            neural crest
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11126)
  AUTHORS   Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JUL-2017) Developmental Biology, Instituto Superior de
            Investigaciones Biologicas (INSIBIO, CONICET), Chacabuco 461, San 
            Miguel de Tucuman, Tucuman T4000ILI, Argentina
REFERENCE   3  (bases 1 to 11126)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11126
                     /organism="Cloning vector Alpha-3000-CRgli2PSM"
                     /mol_type="other DNA"
                     /db_xref="taxon:2052078"
     CDS             9..4391
                     /codon_start=1
                     /gene="Gli2"
                     /product="Gli2"
                     /label=Gli2
                     /note="full length Gli2 protein coding sequence with 6 
                     point mutations in the PKA phosphorilation sites in the 
                     vector alpha-3000 and mutation in the morpholino binding 
                     site"
                     /protein_id="ATQ62676.1"
                     /translation="MEDYVGCVHNSPTLSMISAARGLSPAEVAHEHLKERGIYGLAPPP
                     PPGTTPTEYCHQMAFLASHASPYGELLVQSAAAGNTSHLHDYLTPMDVSRFSSPRVTPR
                     LSRKRALSISPLSDASIDLQTMIRTSPNSLVAYINNSRSSSAASGSYGHLSAGAISPAF
                     SFPIHKPCSLSAALSQQRSLSSSFGHTPLLHPSPTFASRQQGALTSANPAPPSNNSSAP
                     DSVLNKVSSESAVSSTVNQVIHKRSKVKTEEEADSVRFPQPPDHLTDLKEDLDKDECKQ
                     EPEHIYETNCHWDGCSKEFDTQDQLVHHINNDHIHGEKKEFVCRWQDCSREQKPFKAQY
                     MLVVHMRRHTGEKPHKCTFEGCFKAYSRLENLKTHLRSHTGEKPYVCDHEGCNKAFSNA
                     SDRAKHQNRTHSNEKPYICKVPGCTKRYTDPSSLRKHVKTVHGPEAHVTKKHRNDIIQK
                     PSLPKENGDNEASAKLSGREHSDSVSRDQEHCLQTRTIKTEDNMMHQSSPGGQSSCSSE
                     PSPYGNTNNIDSGVDVSLAWQGSLGDLFGLEETSPVVDSTVSSWQRSGRPATPETQRIH
                     SAETGTAEEREIKDNERFLLIYEPNATCQNTRLPTISANGFDVIGVPSSVLINPRAIEL
                     SMNDVTMMNQLNERRDATSSTLSSAYTSRRSAGISPYFSSRRSAETSQFGGRLNNSSSA
                     DSYDPISTDASRRSAEASQHSGLPNLLNLTPAQHYRLKAKYAAATGGPPPTPLPNMDRI
                     GLRNKLSLMDGADFPLPPFRQLPVPRRCADGGGNAGLTPMYPHEIPGNNSRRAADPVRR
                     TAGIDDKPLPRFSRFHSMNSMNTLHPPSLSERRNGGLQHYTCSDGGLHRHVYSPRPPSI
                     SENVAMEAISCDADVPGGDDDLMLPDDVVQYIRSQNREAPEQNLQTEYSSPARNLQSNT
                     KSFHNNTPEQPRAPGAYLSRNFPALAECLGQTANMQDNNMPVQWNEVSSGTVDVSDLPK
                     QQFAAGNLAVVQQKQNFAQYQSFNQAPMQRAHNIMGQGQESVQRNISVNGQRFNYLQQR
                     QQQMSQCQIVSSDFIPQQRYSQSQSMLSSRAMQEGQSQISPSCNNMVERPGVHTHAAPS
                     NTLNHQRLAVHGAPTQGFANNFSVNQDGLHPPNAYTVQPQKNGLEPQQNTLGMSGQAFN
                     HGMIQPRPPAAPHPPNRPRNIHPVHHPPYMRSPHPVSELSPGQQTAEATPKRTSENTDP
                     TPKDNNLLYYSGQIHMYEPNGNFGSGIDCTVRQLPTMPSPGANQVTSTVDSQGLEHPPV
                     IDFDAMMDDGDHSSLMSGTLSPGLLQSFSQTSSRLTTPRNSLTLPSIPAGINNMAIGDM
                     SSMLTTLAEESKFLNLMSYRFKAMEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEED
                     LNEMEQKLISEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDLNSRPLEPLEL"
     gene            9..4391
                     /gene="Gli2"
                     /label=Gli2
     sig_peptide     4107..4361
                     /gene="Gli2"
     CDS             4110..4139
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4149..4178
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4188..4217
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4227..4256
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4266..4295
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4329..4358
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(4386..4403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4408..4542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4641..4659)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5089..5105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5113..5129
                     /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)."
     promoter        complement(5137..5167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5182..5203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5491..6079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6250..7110)
                     /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"
     promoter        complement(7111..7215)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7689..7705
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      8027..11111
                     /regulatory_class="promoter"
     regulatory      10934..10939
                     /regulatory_class="TATA_box"

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