LOCUS       Exported                5079 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCS2+gli3GFP, complete sequence.
ACCESSION   KY435757
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5079)
  AUTHORS   Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Tribulo C, Aybar
            MJ.
  TITLE     The gli3 transcriptional activator is a key effector of Indian 
            hedgehog signaling during Xenopus neural crest development
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5079)
  AUTHORS   Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Tribulo C, Aybar
            MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JAN-2017) Developmental Biology, Instituto Superior de
            Investigaciones Biologicas (INSIBIO, CONICET-UNT), Chacabuco 461, 
            Tucuman, Tucuman T4000ILI, Argentina
REFERENCE   3  (bases 1 to 5079)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: vector nti v. 11
            Sequencing Technology :: Applied Biosystems 3130 Genetic Analyzer 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5079
                     /organism="Cloning vector pCS2+gli3GFP"
                     /mol_type="other DNA"
                     /label=pCS2+
                     /note="pCS2+"
                     /db_xref="taxon:1969590"
     CDS             8..1033
                     /codon_start=1
                     /product="gli3-GFP"
                     /label=gli3-GFP
                     /note="Chimeric protein of gli3 fused to GFP"
                     /protein_id="ARB02202.1"
                     /translation="MEAQSRSTTASEKKKVENSIVKGHSRTEVSEKAVASSTTSNEDES
                     PGQTYHRERRNAIAMQPQGGQGLGKISEEPSTSSEERAMVSKGEELFTGVVPILVELDG
                     DVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMK
                     QHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILG
                     HKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPD
                     NHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGGRSGNSRPLEPLDSAA
                     L"
     CDS             257..973
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(1027..1045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1050..1184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1297..1315)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1336..1352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1360..1376
                     /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(1384..1414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1429..1450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1738..2326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2497..3357)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="Ampicillin resistance"
                     /protein_id="ARB02203.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3358..3462)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3488..3943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      4015..5001
                     /regulatory_class="promoter"
                     /note="simian CMV IE94 enhancer/promoter"
     promoter        4017..5001
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        5036..5054
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"