LOCUS       Exported                7664 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSK485, complete sequence.
ACCESSION   HM640258
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7664)
  AUTHORS   Hartmann T, Dumig M, Jaber BM, Szewczyk E, Olbermann P, Morschhauser
            J, Krappmann S.
  TITLE     Validation of a Self-Excising Marker in the Human Pathogen 
            Aspergillus fumigatus by Employing the {beta}-Rec/six Site-Specific 
            Recombination System
  JOURNAL   Appl. Environ. Microbiol. 76 (18), 6313-6317 (2010)
  PUBMED    20656854
REFERENCE   2  (bases 1 to 7664)
  AUTHORS   Duemig M, Krappmann S.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JUL-2010) YIG3, Research Center for Infectious 
            Diseases, Josef-Scneider-Str. 2/D15, Wuerzburg 97080, Germany
REFERENCE   3  (bases 1 to 7664)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7664
                     /organism="Cloning vector pSK485"
                     /mol_type="other DNA"
                     /label=derived from plasmid pMA-RQ
                     /note="derived from plasmid pMA-RQ"
                     /db_xref="taxon:878333"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_recomb     389..478
                     /label=six site
                     /note="six site"
     regulatory      484..2171
                     /regulatory_class="promoter"
                     /note="derived from Penicillium chrysogenum xylP; similar 
                     to INSD accession M98458.2"
     regulatory      2028..2032
                     /regulatory_class="CAAT_signal"
     regulatory      2095..2099
                     /regulatory_class="TATA_box"
     CDS             2177..2794
                     /codon_start=1
                     /gene="beta"
                     /product="beta-recombinase"
                     /label=beta
                     /note="beta-rec; codon optimized for expression in 
                     Aspergillus fumigatus"
                     /protein_id="ADM43596.1"
                     /translation="MAKIGYARVSSKEQNLDRQLQALQGVSKVFSDKLSGQSVERPQLQ
                     AMLNYIREGDIVVVTELDRLGRNNKELTELMNAIQQKGATLEVLNLPSMNGIEDENLRR
                     LINNLVIELYKYQAESERKRIKERQAQGIEIAKSKGKFKGRQHKFKENDPRLKHAFDLF
                     LNGCSDKEVEEQTGINRRTFRRYRTRYNVTVDQRKNKGKRDS"
     gene            2177..2794
                     /gene="beta"
                     /label=beta
     regulatory      2795..3572
                     /regulatory_class="terminator"
                     /note="derived from Aspergillus nidulans trpC"
     misc_feature    complement(3573..5573)
                     /label=confers resistance to pyrithiamine
                     /note="confers resistance to pyrithiamine"
     CDS             complement(join(3797..4443,4502..4838))
                     /codon_start=1
                     /gene="ptrA"
                     /product="ThiA protein"
                     /label=ptrA
                     /note="putative thiazole synthase; pyrithiamine resistance 
                     protein"
                     /protein_id="ADM43597.1"
                     /translation="MSPPAAIYEPTVAATGLKGKVVVSETVPVEGASQTKLLDHFGGKW
                     DEFKFAPIRESQVSRAMTRRYFEDLDKYAESDVVIVGAGSCGLSTAYVLAKARPDLKIA
                     IVEASVSPGGGAWLGGQLFSAMVMRRPAEVFLNELGVPYEEDANPNYVVVKHASLFTST
                     LMSKVLSFPNVKLFNATAVEDLITRPTENGNPQIAGVVVNWTLVTLHHDDHSCMDPNTI
                     NAPVIISTTGHDGPFGAFCAKRLVSMGSVDKLGGMRGLDMNSAEDAIVKNTREVTKGLI
                     IGGMELSEIDGFNRMGPTFGAMVLSGVKAAEEALKVFDERQRECAE"
     gene            complement(3797..4838)
                     /gene="ptrA"
                     /label=ptrA
     CDS             5236..5247
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_recomb     5583..5672
                     /label=six site
                     /note="six site"
     rep_origin      complement(5861..6528)
                     /direction=LEFT
                     /label=Col E1
                     /note="Col E1"
     rep_origin      complement(5917..6505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6676..7536)
                     /codon_start=1
                     /gene="bla"
                     /product="AmpR"
                     /label=bla
                     /protein_id="ADM43595.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6676..7536)
                     /gene="bla"
                     /label=bla
     promoter        complement(7537..7641)
                     /gene="bla"
                     /label=AmpR promoter