LOCUS       Exported                4223 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pET-SP6p-lacO-GFP-LVA, complete sequence.
ACCESSION   KM229386
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4223)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     A unified design space of synthetic stripe-forming networks
  JOURNAL   Nat Commun 5, 4905 (2014)
  PUBMED    25247316
REFERENCE   2  (bases 1 to 4223)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2014) Centre for Genomic Regulation (CRG), 
            EMBL/CRG Systems Biology Research Unit, Dr. Aiguader 88, Barcelona, 
            Barcelona 08003, Spain
REFERENCE   3  (bases 1 to 4223)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4223
                     /organism="Cloning vector pET-SP6p-lacO-GFP-LVA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1531952"
     CDS             495..686
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    788..930
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1116..1704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1875..2735)
                     /codon_start=1
                     /gene="ampR"
                     /product="ampicillin resistance protein"
                     /label=ampR
                     /note="confers resistance to ampicillin"
                     /protein_id="AIO11022.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1875..2735)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(2736..2840)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2880..2896
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            2909..3846
                     /gene="gfp-LVA"
                     /label=gfp-LVA
     regulatory      2909..2927
                     /regulatory_class="promoter"
                     /gene="gfp-LVA"
                     /note="SP6 promoter"
     promoter        2909..2927
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    2929..2953
                     /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)."
     misc_binding    2931..2951
                     /gene="gfp-LVA"
                     /label=lacI-LVA binding site
                     /bound_moiety="lacI-LVA"
                     /note="lacO"
     RBS             3061..3083
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3091..3846
                     /codon_start=1
                     /gene="gfp-LVA"
                     /product="green fluorescent protein GFP-LVA"
                     /label=gfp-LVA
                     /note="superfolder GFP with degradation tag (LVA)"
                     /protein_id="AIO11021.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKRPAANDENYALVA"
     CDS             3091..3804
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      3954..4206
                     /regulatory_class="terminator"
     terminator      3954..4001
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4010..4096
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4188..4215
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"