LOCUS       Exported                4230 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Synthetic vector pET-T7p(-12T)-SpacerA-GFP-LVA, complete sequence.
ACCESSION   KM261837
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4230)
  AUTHORS   Schaerli Y, Gili M, Isalan M.
  TITLE     A split intein T7 RNA polymerase for transcriptional AND-logic
  JOURNAL   Nucleic Acids Res. (2014) In press
  PUBMED    25262348
REFERENCE   2  (bases 1 to 4230)
  AUTHORS   Schaerli Y, Gili M, Isalan M.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JUL-2014) Centre for Genomic Regulation (CRG), 
            EMBL/CRG Systems Biology Research Unit, Dr. Aiguader 88, Barcelona 
            08003, Spain
REFERENCE   3  (bases 1 to 4230)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4230
                     /organism="synthetic vector pET-T7p(-12T)-SpacerA-GFP-LVA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1550576"
     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="AIS22810.1"
                     /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..3853
                     /gene="GFP-LVA"
                     /label=GFP-LVA
     regulatory      2909..2926
                     /regulatory_class="promoter"
                     /gene="GFP-LVA"
                     /note="T7(-12T) promoter"
     misc_feature    2927..2960
                     /gene="GFP-LVA"
                     /label=spacerA
                     /note="spacerA"
     RBS             3068..3090
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3098..3853
                     /codon_start=1
                     /gene="GFP-LVA"
                     /product="green fluorescent protein (GFP)-LVA"
                     /label=GFP-LVA
                     /note="superfolder GFP with degradation tag (LVA)"
                     /protein_id="AIS22811.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKRPAANDENYALVA"
     CDS             3098..3811
                     /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      3961..4008
                     /regulatory_class="terminator"
     terminator      3961..4008
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4017..4103
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4195..4222
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"