LOCUS       Exported                7062 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pQLinkHD, complete sequence.
ACCESSION   EF025686
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7062)
  AUTHORS   Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K.
  TITLE     Vectors for co-expression of an unrestricted number of proteins
  JOURNAL   Nucleic Acids Res. 35 (6), E43 (2007)
  PUBMED    17311810
REFERENCE   2  (bases 1 to 7062)
  AUTHORS   Buessow K, Scheich C.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-SEP-2006) Vertebrate Genomics, Max Planck Institute 
            for Molecular Genetics, Ihnestr. 63-73, Berlin 14195, Germany
REFERENCE   3  (bases 1 to 7062)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7062
                     /organism="Cloning vector pQLinkHD"
                     /mol_type="other DNA"
                     /db_xref="taxon:405931"
     misc_feature    7..27
                     /label=LIC cassette with PacI restriction site
                     /note="LIC cassette with PacI restriction site"
     promoter        36..80
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    56..72
                     /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)."
     regulatory      56..61
                     /regulatory_class="minus_35_signal"
     regulatory      74..79
                     /regulatory_class="minus_10_signal"
     protein_bind    88..104
                     /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)."
     RBS             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      127..133
                     /regulatory_class="ribosome_binding_site"
     misc_feature    141
                     /note="translation start site for inserts cloned into the 
                     multiple cloning site"
     CDS             147..167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     misc_feature    180..337
                     /label=attR1 recombination site
                     /note="attR1 recombination site"
     protein_bind    180..304
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    647..2453
                     /note="multiple cloning site; includes BamHI, NotI and 
                     HindIII restriction sites"
     CDS             complement(733..1038)
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /note="encodes a cytotoxic protein that is a potent poison 
                     of DNA gyrase"
                     /protein_id="ABJ15763.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1383..2042)
                     /codon_start=1
                     /gene="cat-1"
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label=cat-1
                     /protein_id="ABJ15764.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(1383..2042)
                     /gene="cat-1"
                     /label=cat-1
     promoter        complement(2043..2145)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_feature    2290..2364
                     /label=attR2 recombination site
                     /note="attR2 recombination site"
     protein_bind    complement(2323..2447)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2469..2563
                     /regulatory_class="terminator"
                     /note="lambda t0 transcriptional terminator"
     terminator      2469..2563
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    2568..2617
                     /label=LIC cassette with SwaI, PacI restriction sites
                     /note="LIC cassette with SwaI, PacI restriction sites"
     CDS             2657..3316
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      3375..3472
                     /regulatory_class="terminator"
                     /note="rrnB T1 transcriptional terminator"
     terminator      3381..3467
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3560..4642)
                     /codon_start=1
                     /gene="lacI"
                     /allele="Q"
                     /product="lac repressor"
                     /label=lacI
                     /note="overproducing allele; similar to lac repressor 
                     encoded by GenBank Accession Number U78872"
                     /protein_id="ABJ15762.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3560..4642)
                     /gene="lacI"
                     /allele="Q"
                     /label=lacI
     promoter        complement(4643..4720)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    4912..5052
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      5236
                     /label=ColE1 ori
                     /note="ColE1 ori"
     rep_origin      complement(5238..5826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5997..6857)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /note="penicillinase; similar to the protein encoded by 
                     GenBank Accession Number AF074376"
                     /protein_id="ABJ15761.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5997..6857)
                     /gene="bla"
                     /label=bla
     promoter        complement(6858..6962)
                     /gene="bla"
                     /label=AmpR promoter