LOCUS       Exported                5989 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Bacterial expression vector pTrcmelA, complete sequence.
ACCESSION   LT726808
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5989)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5989)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 5989)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5989
                     /organism="Bacterial expression vector pTrcmelA"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 9152."
                     /db_xref="taxon:1944709"
     CDS             complement(41..1870)
                     /codon_start=1
                     /note="unnamed protein product; melA"
                     /protein_id="SJL86559.1"
                     /translation="MAWLVGKPSLERSWNAILSFPESGFQLECRNTIGSSVFSSHFTLH
                     FRVARRLLHFSCRRFTETQKEPTQALWWCELPTAPAPRRRGTGLKAALILAKDNSNPRE
                     SKMSITRRHVIVQGGVIAAGLLASGLPGTKAFAQIPSIPWRRSLQGLAWNDPIIETYRD
                     AVRLLNALPASDKFNWVNLSKIHGSGDVVKYCPHGNWYFLPWHRAYTAMYERIVRHVTK
                     NNDFAMPFWDWTDNPYLPEVFTMQKTPDGKDNPLYVSSRTWPITQPMPDNIVGPQVLNT
                     ILTAKPYEVFGTTRPEGQNSLDPSWVTTSSGTQGALEYTPHNQVHNNIGGWMPEMSSPR
                     DPIFFMHHCNIDRIWATWNLRNANSTDRLWADMPFTDNFYDVDGNFWSPKVSDLYVPEE
                     LGYNYGFRTYFKVAAASAKTLALNDKLTSVIAATATDAAIAGVTTTSTDNSKAATENVP
                     LSLPIKIPAGALQEIVRQPPLPSGMDTMDFGAAQEQAASAPRVLAFLRDVEITSASTTS
                     VRVFLGKNDLKADTPVTDPHYVGSFAVLGHDGDHHRKPSFVLDLTDAIQRVYGGRGQTD
                     GEAIDLQLIPVGSGAGKPGAVEPAKLEIAIVSA"
     misc_feature    complement(1874..1909)
                     /label=5 UTR of lacZ, incl. RBS
                     /note="5 UTR of lacZ, incl. RBS"
     protein_bind    1891..1907
                     /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      complement(1910..1953)
                     /regulatory_class="promoter"
                     /note="trc promoter"
     promoter        complement(1915..1944)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(2176..3258)
                     /codon_start=1
                     /note="unnamed protein product; lacI"
                     /protein_id="SJL86560.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     regulatory      complement(3259..3336)
                     /regulatory_class="promoter"
                     /note="lacIq promoter, incl. RBS"
     promoter        complement(3259..3336)
                     /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    3522..3662
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3664..4066)
                     /direction=LEFT
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      complement(3848..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4607..5467)
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL86561.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5468..5559)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(5579..5606)
                     /regulatory_class="terminator"
                     /note="T2 terminator"
     terminator      5579..5606
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5698..5784
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      5738..5781
                     /regulatory_class="terminator"
                     /note="T1 terminator"