LOCUS       Exported                4360 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTM30, complete sequence.
ACCESSION   AF023620
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4360)
  AUTHORS   Morrison TB, Parkinson JS.
  TITLE     Liberation of an interaction domain from the phosphotransfer region 
            of CheA, a signaling kinase of Escherichia coli
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 91 (12), 5485-5489 (1994)
  PUBMED    8202513
REFERENCE   2  (bases 1 to 4360)
  AUTHORS   Morrison TB.
  TITLE     pTM30 expression vector for randomly coding DNA fragments
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 4360)
  AUTHORS   Morrison TB, Parkinson JS.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-SEP-1997) Pathology, University of Utah, School of 
            Medicine 5B210, Salt Lake City, UT 84132, USA
REFERENCE   4  (bases 1 to 4360)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4360
                     /organism="Cloning vector pTM30"
                     /mol_type="other DNA"
                     /note="derived in part from pBR322; pUC119"
                     /db_xref="taxon:67588"
     regulatory      1..31
                     /regulatory_class="promoter"
                     /note="Ptac"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     promoter        2..30
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..59
                     /function="LacI operator"
                     /label=LacI binding site
                     /bound_moiety="LacI"
     protein_bind    38..54
                     /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      60..70
                     /regulatory_class="ribosome_binding_site"
                     /gene="cheY"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     gene            60..70
                     /gene="cheY"
                     /label=cheY
     misc_feature    74..102
                     /label=Region: cloning site
                     /note="Region: cloning site"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     regulatory      103..113
                     /regulatory_class="terminator"
                     /note="ochre translation terminators in all reading frames"
                     /experiment="experimental evidence, no additional details 
                     recorded"
     rep_origin      187..642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        850..927
                     /gene="lacI"
                     /label=lacI promoter
     CDS             928..2010
                     /codon_start=1
                     /gene="lacI"
                     /allele="q"
                     /product="repressor LacI-q"
                     /function="suppresses Ptac transcription"
                     /label=lacI
                     /protein_id="AAB83985.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGAGISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            928..2010
                     /gene="lacI"
                     /allele="q"
                     /label=lacI
                     /note="EcoRV site removed by mutagenesis"
     variation       1727
                     /gene="lacI"
                     /replace="a"
                     /label=destroyed EcoRV site
                     /note="destroyed EcoRV site"
     protein_bind    2023..2044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2170..2274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2275..3135
                     /codon_start=1
                     /gene="blaC"
                     /product="beta lactamase"
                     /label=blaC
                     /protein_id="AAB83984.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPPAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2275..3135
                     /gene="blaC"
                     /label=blaC
     rep_origin      3306..3894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      3894
     misc_feature    4080..4220
                     /label=bom
                     /note="basis of mobility region from pBR322"