LOCUS       Exported                5434 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWSK29, complete sequence.
ACCESSION   AF016889
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5434)
  AUTHORS   Wang RF, Kushner SR.
  TITLE     Construction of versatile low-copy-number vectors for cloning, 
            sequencing and gene expression in Escherichia coli
  JOURNAL   Gene 100, 195-199 (1991)
  PUBMED    2055470
REFERENCE   2  (bases 1 to 5434)
  AUTHORS   Maples VF, Kushner SR.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-AUG-1997) Genetics, University of Georgia, Life 
            Sciences Bldg., Athens, GA 30602, USA
REFERENCE   3  (bases 1 to 5434)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5434
                     /organism="Cloning vector pWSK29"
                     /host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:65736"
     misc_feature    1..2293
                     /label=originates from pBluescript II SK(+)
                     /note="originates from pBluescript II SK(+)"
     CDS             complement(144..1004)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAB81041.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(144..1004)
                     /gene="bla"
                     /label=bla
     misc_difference 519
                     /replace="a"
                     /label=pBluescript II SK(+)
                     /note="pBluescript II SK(+)"
     promoter        complement(1005..1109)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1135..1590)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1373..1948)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     1732..1748
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1758..1776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1785..1892
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1802..1818
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1852..1868)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1905..1923)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1944..1960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1968..1984
                     /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(1992..2022)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2037..2058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2351..5270
                     /label=pLg339
                     /note="pLg339"
     misc_feature    2351..5270
                     /label=similar to pSC101
                     /note="similar to pSC101"
     misc_difference 2641
                     /label=pSC101
                     /note="pSC101"
     misc_difference 2830
                     /replace="c"
                     /label=pSC101
                     /note="pSC101"
     rep_origin      3337..3559
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             3607..4557
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiation protein"
                     /label=rep
                     /protein_id="AAB81042.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     gene            3607..4557
                     /gene="rep"
                     /label=rep
                     /note="pLG339 replication protein"
     misc_difference 4376
                     /replace="g"
                     /label=pSC101
                     /note="pSC101"
     misc_difference 4471
                     /replace="c"
                     /label=pSC101
                     /note="pSC101"
     misc_difference 4633
                     /replace="g"
                     /label=pSC101
                     /note="pSC101"
     misc_difference 4723
                     /replace="t"
                     /label=pSC101
                     /note="pSC101"
     misc_difference 4997..4999
                     /label=pSC101
                     /note="pSC101"
     misc_feature    5271..5431
                     /note="originates from pLG339; contains portion of tn903 
                     (kanamycin resistance transposon)"