LOCUS       Exported                8438 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCS2+MT-CRgli2, complete sequence.
ACCESSION   MF444683
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  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  JOURNAL   Unpublished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 8438)
  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 8438)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8438
                     /organism="Cloning vector pCS2+MT-CRgli2"
                     /mol_type="other DNA"
                     /db_xref="taxon:2052079"
     misc_binding    7..31
                     /label=mutated morpholino binding site
                     /bound_moiety="mutated morpholino binding site"
     CDS             10..4392
                     /codon_start=1
                     /gene="Gli2"
                     /product="Gli2"
                     /label=Gli2
                     /note="full length Gli2 protein coding sequence with a 
                     mutation in the morpholino binding site"
                     /protein_id="ATQ62674.1"
                     /translation="MEDYVGCVHNSPTLSMISAARGLSPAEVAHEHLKERGIYGLAPPP
                     PPGTTPTEYCHQMAFLASHASPYGELLVQSAAAGNTSHLHDYLTPMDVSRFSSPRVTPR
                     LSRKRALSISPLSDASIDLQTMIRTSPNSLVAYINNSRSSSAASGSYGHLSAGAISPAF
                     SFPIHKPCSLSAALSQQRSLSSSFGHTPLLHPSPTFASRQQGALTSANPAPPSNNSSAP
                     DSVLNKVSSESAVSSTVNQVIHKRSKVKTEEEADSVRFPQPPDHLTDLKEDLDKDECKQ
                     EPEHIYETNCHWDGCSKEFDTQDQLVHHINNDHIHGEKKEFVCRWQDCSREQKPFKAQY
                     MLVVHMRRHTGEKPHKCTFEGCFKAYSRLENLKTHLRSHTGEKPYVCDHEGCNKAFSNA
                     SDRAKHQNRTHSNEKPYICKVPGCTKRYTDPSSLRKHVKTVHGPEAHVTKKHRNDIIQK
                     PSLPKENGDNEASAKLSGREHSDSVSRDQEHCLQTRTIKTEDNMMHQSSPGGQSSCSSE
                     PSPYGNTNNIDSGVDVSLAWQGSLGDLFGLEETSPVVDSTVSSWQRSGRPATPETQRIH
                     SAETGTAEEREIKDNERFLLIYEPNATCQNTRLPTISANGFDVIGVPSSVLINPRAIEL
                     SMNDVTMMNQLNERRDSTSSTLSSAYTSRRSSGISPYFSSRRSSETSQFGGRLNNSSSA
                     DSYDPISTDASRRSSEASQHSGLPNLLNLTPAQHYRLKAKYAAATGGPPPTPLPNMDRI
                     GLRNKLSLMDGADFPLPPFRQLPVPRRCSDGGGNAGLTPMYPHEIPGNNSRRASDPVRR
                     TAGIDDKPLPRFSRFHSMNSMNTLHPPSLSERRNGGLQHYTCSDGGLHRHVYSPRPPSI
                     SENVAMEAISCDADVPGGDDDLMLPDDVVQYIRSQNREAPEQNLQTEYSSPARNLQSNT
                     KSFHNNTPEQPRAPGAYLSRNFPALAECLGQTANMQDNNMPVQWNEVSSGTVDVSDLPK
                     QQFAAGNLAVVQQKQNFAQYQSFNQAPMQRAHNIMGQGQESVQRNISVNGQRFNYLQQR
                     QQQMSQCQIVSSDFIPQQRYSQSQSMLSSRAMQEGQSQISPSCNNMVERPGVHTHAAPS
                     NTLNHQRLAVHGAPTQGFANNFSVNQDGLHPPNAYTVQPQKNGLEPQQNTLGMSGQAFN
                     HGMIQPRPPAAPHPPNRPRNIHPVHHPPYMRSPHPVSELSPGQQTAEATPKRTSENTDP
                     TPKDNNLLYYSGQIHMYEPNGNFGSGIDCTVRQLPTMPSPGANQVTSTVDSQGLEHPPV
                     IDFDAMMDDGDHSSLMSGTLSPGLLQSFSQTSSRLTTPRNSLTLPSIPAGINNMAIGDM
                     SSMLTTLAEESKFLNLMSYRFKAMEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEED
                     LNEMEQKLISEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDLNSRPLEPLEL"
     gene            10..4392
                     /gene="Gli2"
                     /label=Gli2
     CDS             4111..4140
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4150..4179
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4189..4218
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4228..4257
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4267..4296
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4330..4359
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(4387..4404)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(4388..4404)
                     /regulatory_class="promoter"
     polyA_signal    4409..4543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4656..4674)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4695..4711)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4719..4735
                     /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(4743..4773)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4788..4809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5097..5685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5856..6716)
                     /codon_start=1
                     /gene="Amp"
                     /product="ampicilin resistance protein"
                     /label=Amp
                     /protein_id="ATQ62675.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5856..6716)
                     /gene="Amp"
                     /label=Amp
     promoter        complement(6717..6821)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6847..7302)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7376..8360
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        8395..8413
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      8395..8411
                     /regulatory_class="promoter"