LOCUS       Exported                5165 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pOKE104, complete sequence.
ACCESSION   FJ599664
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5165)
  AUTHORS   Low W, Jedd G.
  TITLE     An improved plasmid for transformation of Neurospora crassa using 
            the pan-2 gene as a selectable marker
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5165)
  AUTHORS   Low W, Jedd G.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-2008) Temasek Life Sciences Laboratory, 1 Research
            Link National University of Singapore 117604, Singapore
REFERENCE   3  (bases 1 to 5165)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5165
                     /organism="Transformation vector pOKE104"
                     /mol_type="other DNA"
                     /label=used in Neurospora crassa
                     /note="used in Neurospora crassa"
                     /db_xref="taxon:588564"
     misc_feature    5..61
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    6..56
                     /label=polylinker
                     /note="polylinker"
     promoter        complement(68..86)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(104..120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    128..144
                     /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)."
     promoter        complement(152..182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    197..218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(506..1094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1265..2125)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="ACM43516.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1265..2125)
                     /gene="bla"
                     /label=bla
     promoter        complement(2126..2230)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2889..2912
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             join(2989..3255,3473..4135)
                     /codon_start=1
                     /gene="pan-2"
                     /product="3-methyl-2-oxobutanoate hydroxymethyltransferase"
                     /label=pan-2
                     /note="pantothenic acid-2"
                     /protein_id="ACM43517.1"
                     /translation="MGLPPTNPRKKVTLNTLQSMYKKGEKITMITAHDFPSAHVAEHAG
                     MDMILVGDSLAMVALGMEDTSEVLVEEMLLHCRSVARATSRAFTVGDLPMGSYEISPQQ
                     ALSTAIRFIKEGRVSAVKLEGGAEMAPTIRAITTAGIPVLAHIGLTPQRQNALGGFRVQ
                     GKSKESAKRLLRDALAVQEAGAFAVVVEAVPQEVAAFITGLLKIPTIGIGAGNGTSGQV
                     LVQADMTGNFPPGRFLPKFVKKYGDVWGEALRAIETYRDEVKGGSYPAEEHTYPMAKEE
                     VEAFVGDVRRLMNANASAKAEGEGEKDA"
     gene            2989..4135
                     /gene="pan-2"
                     /label=pan-2
     rep_origin      4530..4985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4977..108)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL"
     primer_bind     5126..5142
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5149..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"