LOCUS       Exported                5431 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p1enVT-lacI, complete sequence.
ACCESSION   KX986156
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  AUTHORS   Boehm CR, Grant PK, Haseloff J.
  TITLE     Programmed emergence of a domain of gene expression in a bacterial 
            population
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5431)
  AUTHORS   Boehm CR.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-OCT-2016) Plant Sciences, University of Cambridge, 
            Downing Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom
REFERENCE   3  (bases 1 to 5431)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5431
                     /organism="Cloning vector p1enVT-lacI"
                     /mol_type="other DNA"
                     /db_xref="taxon:2049404"
     rep_origin      complement(49..637)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    818..902
                     /note="PT7 and transl. enh. RBS from pET3a (Rosenberg et 
                     al. 1987)"
     promoter        825..843
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             875..897
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             909..1628
                     /codon_start=1
                     /product="mVenus"
                     /label=mVenus
                     /protein_id="ATP60589.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1640..1768
                     /regulatory_class="terminator"
                     /note="BBa_B0015"
     terminator      1648..1719
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1735..1762
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      1769..1803
                     /regulatory_class="promoter"
                     /note="PJ23101*"
     misc_feature    1812..3119
                     /note="lacI_generator cassette; BBa_P0412"
     RBS             1812..1823
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1839..2918
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             2919..2951
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      2999..3070
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3086..3113
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3120..3140
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3166..4023)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4024..4128)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      4218..4261
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4264..4285
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     regulatory      complement(4286..4414)
                     /regulatory_class="terminator"
                     /note="BBa_B0015"
     terminator      4292..4319
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      4335..4406
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4426..5145)
                     /codon_start=1
                     /product="mTurquoise2"
                     /label=mTurquoise2
                     /protein_id="ATP60590.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      complement(5146..5163)
                     /regulatory_class="ribosome_binding_site"
                     /note="BBa_B0034"
     RBS             5152..5163
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      complement(5172..5206)
                     /regulatory_class="promoter"
                     /note="pJ23101"
     misc_feature    5207..5227
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5228..5285
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."