LOCUS       Exported               11145 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector Alpha3000-MT-gli2-6xHIS, complete sequence.
ACCESSION   MF435043
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11145)
  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  JOURNAL   Unpublished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 11145)
  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 11145)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11145
                     /organism="Cloning vector Alpha3000-MT-gli2-6xHIS"
                     /mol_type="other DNA"
                     /db_xref="taxon:2052062"
     CDS             21..4397
                     /codon_start=1
                     /gene="Gli2"
                     /product="Gli2"
                     /label=Gli2
                     /note="full length coding sequence of Xenopus laevis Gli2 
                     protein"
                     /protein_id="ATQ62671.1"
                     /translation="MEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEEDLNEMEQKLI
                     SEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDLNSRPLDVEHYLRSVHNSPTL
                     SMISAARGLSPAEVAHEHLKERGIYGLAPPPPGTTPTEYCHQMAFLASHASPYGELLVQ
                     SAAAGNTSHLHDYLTPMDVSRFSSPRVTPRLSRKRALSISPLSDASIDLQTMIRTSPNS
                     LVAYINNSRSSSAASGSYGHLSAGAISPAFSFPIHKPCSLSAALSQQRSLSSSFGHTPL
                     LHPSPTFASRQQGALTSANPAPPSNNSSAPDSVLNKVSSESAVSSTVNQVIHKRSKVKT
                     EEEADSVRFPQPPDHLTDLKEDLDKDECKQEPEHIYETNCHWDGCSKEFDTQDQLVHHI
                     NNDHIHGEKKEFVCRWQDCSREQKPFKAQYMLVVHMRRHTGEKPHKCTFEGCFKAYSRL
                     ENLKTHLRSHTGEKPYVCDHEGCNKAFSNASDRAKHQNRTHSNEKPYICKVPGCTKRYT
                     DPSSLRKHVKTVHGPEAHVTKKHRNDIIQKPSLPKENGDNEASAKLSGREHSDSVSRDQ
                     EHCLQTRTIKTEDNMMHQSSPGGQSSCSSEPSPYGNTNNIDSGVDVSLAWQGSLGDLFG
                     LEETSPVVDSTVSSWQRSGRPATPETQRIHSAETGTAEEREIKDNERFLLIYEPNATCQ
                     NTRLPTISANGFDVIGVPSSVLINPRAIELSMNDVTMMNQLNERRDSTSSTLSSAYTSR
                     RSSGISPYFSSRRSSETSQFGGRLNNSSSADSYDPISTDASRRSSEASQHSGLPNLLNL
                     TPAQHYRLKAKYAAATGGPPPTPLPNMDRIGLRNKLSLMDGADFPLPPFRQLPVPRRCS
                     DGGGNAGLTPMYPHEIPGNNSRRASDPVRRTAGIDDKPLPRFSRFHSMNSMNTLHPPSL
                     SERRNGGLQHYTCSDGGLHRHVYSPRPPSISENVAMEAISCDADVPGGDDDLMLPDDVV
                     QYIRSQNREAPEQNLQTEYSSPARNLQSNTKSFHNNTPEQPRAPGAYLSRNFPALAECL
                     GQTANMQDNNMPVQWNEVSSGTVDVSDLPKQQFAAGNLAVVQQKQNFAQYQSFNQAPMQ
                     RAHNIMGQGQESVQRNISVNGQRFNYLQQRQQQMSQCQIVSSDFIPQQRYSQSQSMLSS
                     RAMQEGQSQISPSCNNMVERPGVHTHAAPSNTLNHQRLAVHGAPTQGFANNFSVNQDGL
                     HPPNAYTVQPQKNGLEPQQNTLGMSGQAFNHGMIQPRPPAAPHPPNRPRNIHPVHHPPY
                     MRSPHPVSELSPGQQTAEATPKRTSENTDPTPKDNNLLYYSGQIHMYEPNGNFGSGIDC
                     TVRQLPTMPSPGANQVTSTVDSQGLEHPPVIDFDAMMDDGDHSSLMSGTLSPGLLQSFS
                     QTSSRLTTPRNSLTLPSIPAGINNMAIGDMSSMLTTLAEESKFLAIMQLQHHHHHH"
     gene            21..4397
                     /gene="Gli2"
                     /label=Gli2
     sig_peptide     21..275
                     /gene="Gli2"
     CDS             24..53
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             63..92
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             102..131
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             141..170
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             180..209
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             243..272
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     sig_peptide     4377..4394
                     /gene="Gli2"
     CDS             4377..4394
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(4405..4422)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4427..4622
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    4427..4561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4660..4678)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5108..5124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5132..5148
                     /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(5156..5186)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5201..5222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5510..6098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6269..7129)
                     /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(7130..7234)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7708..7724
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      8046..11130
                     /regulatory_class="promoter"
     regulatory      10953..10958
                     /regulatory_class="TATA_box"