LOCUS       Exported                6837 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector Tmp-pSTB205, complete sequence.
ACCESSION   MF509189
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6837)
  AUTHORS   Martinez R.
  TITLE     Cloning challenging DNA sequences: a different approach with linear 
            vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6837)
  AUTHORS   Martinez R.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-JUL-2017) Plant Developmental Biology, Max Planck 
            Institute For Plant Breeding Research, Carl-von-Linne Weg 10, 
            Cologne, North Rhine-Westphalia 50829, Germany
REFERENCE   3  (bases 1 to 6837)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6837
                     /organism="Cloning vector Tmp-pSTB205"
                     /mol_type="other DNA"
                     /db_xref="taxon:2026119"
     regulatory      73..100
                     /regulatory_class="other"
                     /note="rrnB T2"
     terminator      73..100
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      192..278
                     /regulatory_class="other"
                     /note="rrnB T1"
     terminator      192..278
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    331..430
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    331..404
                     /label=attP1
                     /note="attP1"
     misc_feature    405..432
                     /label=attM1 adapter
                     /note="attM1 adapter"
     CDS             complement(828..1133)
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /note="a bacterial toxin that poisons DNA gyrase"
                     /protein_id="AST51818.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1475..2134)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /function="confers resistance to chloramphenicol"
                     /label=chloramphenicol acetyltransferase
                     /protein_id="AST51819.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2135..2237)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_feature    2382..2406
                     /label=attM2 adapter
                     /note="attM2 adapter"
     protein_bind    complement(2383..2482)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2407..2482
                     /label=attP2
                     /note="attP2"
     misc_feature    join(2413,2483..2484)
                     /label=opens ORF to stb
                     /note="opens ORF to stb"
     misc_feature    2485..5112
                     /label=stb
                     /note="stb"
     CDS             join(2485..3201,3232..3542,3805..3947,4024..4631)
                     /codon_start=1
                     /product="Venus"
                     /label=Venus
                     /protein_id="AST51820.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKELLMCSGQAESGGSSSTESSSLSGGLRFGQKIYFEDGSGSRS
                     KNRVNTVRKSSTTARCQVEGCRMDLSNVKAYYSRHKVCCIHSKSSKVIVSGLHQRFCQQ
                     CSRFHQLSEFDLEKRSCRRRLACHNERRRKPQPTTALFTSHYSRIAPSLYGNPNAAMIK
                     SVLGDPTAWSTARSVMQRPGPWQINPVRETHPHMNVLSHGSSSFTTCPEMINNNSTDSS
                     CALSLLSNSYPIHQQQLQTPTNTWRPSSGFDSMISFSDKVTMAQPPPISTHQPPISTHQ
                     QYLSQTWEVIAGEKSNSHYMSPVSQISEPADFQISNGTTMGGFELYLHQQVLKQYMEPE
                     NTRAYDSSPQHFNWSL"
     CDS             2485..3201
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             5235..6044
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /function="confers resistance to kanamycin"
                     /label=aminoglycoside phosphotransferase
                     /protein_id="AST51821.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      6187..6775
                     /direction=RIGHT
                     /label=pUC origin of replication
                     /note="pUC origin of replication"