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  JOURNAL   Unpublished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"