LOCUS       Exported                3659 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning and transformation vector pUC-3GLA, complete sequence.
ACCESSION   KM977568
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3659)
  AUTHORS   Haussmann IU, Brauer U, Ustaoglu P, Hemani Y, Soller M.
  TITLE     Plasmid based gap-repair recombineering for Drosophila transgenesis 
            and the analysis of alternative pre-mRNA processing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3659)
  AUTHORS   Haussmann IU, Brauer U, Ustaoglu P, Hemani Y, Soller M.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2014) School of Bioscience, University of 
            Birmingham, Edgbaston, Birmingham, West Midlands B15 2TT, United 
            Kingdom
REFERENCE   3  (bases 1 to 3659)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3659
                     /organism="Cloning and transformation vector pUC-3GLA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1681183"
     misc_recomb     1..280
                     /note="attB site; bacterial attachment site"
     protein_bind    188..257
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     promoter        625..729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             730..1590
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AKQ12673.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            730..1590
                     /gene="bla"
                     /label=bla
     rep_origin      1761..2349
     misc_feature    2370..2461
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_recomb     2466..2503
                     /label=loxP site for cre mediated recombination
                     /note="loxP site for cre mediated recombination"
     protein_bind    complement(2466..2499)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2504..2740
                     /label=3xP3-hsp70
                     /note="3xP3-hsp70"
     regulatory      2735..2744
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2741..3505
                     /codon_start=1
                     /gene="gfp"
                     /product="green flourescent protein"
                     /label=gfp
                     /note="GFP"
                     /protein_id="AKQ12674.1"
                     /translation="MVSSASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGK
                     LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFF
                     KDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN
                     GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM
                     VLLEFVTAAGITLGMDELYKYSDLCKLVEFPI"
     gene            2741..3505
                     /gene="gfp"
                     /label=gfp
     CDS             2756..3466
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     misc_feature    3503..3602
                     /note="ewg PA1; polyadenylation site 1 of the Drosophila 
                     erect wing gene"
     protein_bind    complement(3614..3647)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     3641..3647
                     /label=loxP site for cre mediated recombination
                     /note="loxP site for cre mediated recombination"