LOCUS       Exported                8114 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCS2+MT-gli2act, complete sequence.
ACCESSION   MF435039
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8114)
  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  JOURNAL   Unpublished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 8114)
  AUTHORS   Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-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 8114)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8114
                     /organism="Cloning vector pCS2+MT-gli2act"
                     /mol_type="other DNA"
                     /db_xref="taxon:2052065"
     CDS             6..4067
                     /codon_start=1
                     /gene="Gli2"
                     /product="Gli2act"
                     /label=Gli2
                     /note="activator form of Gli2 protein"
                     /protein_id="ATQ62666.1"
                     /translation="MGGGALSISPLSDASIDLQTMIRTSPNSLVAYINNSRSSSAASGS
                     YGHLSAGAISPAFSFPIHKPCSLSAALSQQRSLSSSFGHTPLLHPSPTFASRQQGALTS
                     ANPAPPSNNSSAPDSVLNKVSSESAVSSTVNQVIHKRSKVKTEEEADSVRFPQPPDHLT
                     DLKEDLDKDECKQEPEHIYETNCHWDGCSKEFDTQDQLVHHINNDHIHGEKKEFVCRWQ
                     DCSREQKPFKAQYMLVVHMRRHTGEKPHKCTFEGCFKAYSRLENLKTHLRSHTGEKPYV
                     CDHEGCNKAFSNASDRAKHQNRTHSNEKPYICKVPGCTKRYTDPSSLRKHVKTVHGPEA
                     HVTKKHRNDIIQKPSLPKENGDNEASAKLSGREHSDSVSRDQEHCLQTRTIKTEDNMMH
                     QSSPGGQSSCSSEPSPYGNTNNIDSGVDVSLAWQGSLGDLFGLEETSPVVDSTVSSWQR
                     SGRPATPETQRIHSAETGTAEEREIKDNERFLLIYEPNATCQNTRLPTISANGFDVIGV
                     PSSVLINPRAIELSMNDVTMMNQLNERRDSTSSTLSSAYTSRRSSGISPYFSSRRSSET
                     SQFGGRLNNSSSADSYDPISTDASRRSSEASQHSGLPNLLNLTPAQHYRLKAKYAAATG
                     GPPPTPLPNMDRIGLRNKLSLMDGADFPLPPFRQLPVPRRCSDGGGNAGLTPMYPHEIP
                     GNNSRRASDPVRRTAGIDDKPLPRFSRFHSMNSMNTLHPPSLSERRNGGLQHYTCSDGG
                     LHRHVYSPRPPSISENVAMEAISCDADVPGGDDDLMLPDDVVQYIRSQNREAPEQNLQT
                     EYSSPARNLQSNTKSFHNNTPEQPRAPGAYLSRNFPALAECLGQTANMQDNNMPVQWNE
                     VSSGTVDVSDLPKQQFAAGNLAVVQQKQNFAQYQSFNQAPMQRAHNIMGQGQESVQRNI
                     SVNGQRFNYLQQRQQQMSQCQIVSSDFIPQQRYSQSQSMLSSRAMQEGQSQISPSCNNM
                     VERPGVHTHAAPSNTLNHQRLAVHGAPTQGFANNFSVNQDGLHPPNAYTVQPQKNGLEP
                     QQNTLGMSGQAFNHGMIQPRPPAAPHPPNRPRNIHPVHHPPYMRSPHPVSELSPGQQTA
                     EATPKRTSENTDPTPKDNNLLYYSGQIHMYEPNGNFGSGIDCTVRQLPTMPSPGANQVT
                     STVDSQGLEHPPVIDFDAMMDDGDHSSLMSGTLSPGLLQSFSQTSSRLTTPRNSLTLPS
                     IPAGINNMAIGDMSSMLTTLAEESKFLNLNRFKAMEQKLISEEDLNEMEQKLISEEDLN
                     EMEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDL
                     NSRPLEPLEL"
     gene            6..4067
                     /gene="Gli2"
                     /label=Gli2
     sig_peptide     3783..4037
                     /gene="Gli2"
     CDS             3786..3815
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3825..3854
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3864..3893
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3903..3932
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3942..3971
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4005..4034
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(4062..4079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(4063..4079)
                     /regulatory_class="promoter"
     regulatory      4084..4279
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    4084..4218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4331..4349)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4370..4386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4394..4410
                     /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(4418..4448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4463..4484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4772..5360)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5531..6391)
                     /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(6392..6496)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6522..6977)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7051..8035
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     regulatory      7094..8035
                     /regulatory_class="promoter"
     promoter        8070..8088
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      8070..8086
                     /regulatory_class="promoter"