LOCUS       Exported                3293 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning and transformation vector pOT2-3GLA, complete sequence.
ACCESSION   KM977569
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3293)
  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  JOURNAL   Unpublished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 3293)
  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 3293)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: DNASTAR Lasergene SeqBuilder v. 7.2.1 
            Sequencing Technology :: not fully sequenced, assembled from
                                     pieces, vector fully functional 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3293
                     /organism="Cloning and transformation vector pOT2-3GLA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1681182"
     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        complement(387..405)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(470..1129)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="AKQ12675.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(470..1129)
                     /gene="cat"
                     /label=cat
     promoter        complement(1166..1194)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      1241..1848
     rep_origin      complement(1256..1844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      1948..1966
                     /regulatory_class="promoter"
                     /note="T7"
     promoter        1948..1966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2004..2095
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_recomb     2099..2137
                     /label=loxP site for cre mediated recombination
                     /note="loxP site for cre mediated recombination"
     protein_bind    complement(2100..2133)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2138..2374
                     /label=3xP3-hsp70
                     /note="3xP3-hsp70"
     regulatory      2369..2378
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2375..3139
                     /codon_start=1
                     /gene="gfp"
                     /product="green flourescent protein"
                     /label=gfp
                     /note="GFP"
                     /protein_id="AKQ12676.1"
                     /translation="MVSSASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGK
                     LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFF
                     KDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN
                     GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHM
                     VLLEFVTAAGITLGMDELYKYSDLCKLVEFPI"
     gene            2375..3139
                     /gene="gfp"
                     /label=gfp
     CDS             2390..3100
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     misc_feature    3137..3236
                     /note="ewg PA1; polyadenylation site 1 of the Drosophila 
                     erect wing gene"
     misc_recomb     3248..3281
                     /label=loxP site for cre mediated recombination
                     /note="loxP site for cre mediated recombination"
     protein_bind    complement(3248..3281)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."