LOCUS       Exported                5514 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTVP1GFP, complete sequence.
ACCESSION   KM242650
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5514)
  AUTHORS   Garcia-Fruitos E, Gonzalez-Montalban N, Morell M, Vera A, Ferraz RM,
            Aris A, Ventura S, Villaverde A.
  TITLE     Aggregation as bacterial inclusion bodies does not imply 
            inactivation of enzymes and fluorescent proteins
  JOURNAL   Microb. Cell Fact. 4, 27 (2005)
  PUBMED    16156893
REFERENCE   2  (bases 1 to 5514)
  AUTHORS   Garcia-Fruitos E.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2014) Institut de Biotecnologia i de Biomedicina, 
            CIBER-BBN, Autonomous University of Barcelona, Campus Bellaterra, 
            Bellaterra 08193, Spain
REFERENCE   3  (bases 1 to 5514)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5514
                     /organism="Cloning vector pTVP1GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1559749"
     promoter        193..222
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     CDS             267..1622
                     /codon_start=1
                     /product="VP1/green fluorescent fusion protein"
                     /label=VP1/green fluorescent fusion protein
                     /note="VP1/GFP; derived from Foot-and-mouth disease virus 
                     and Aequorea victoria"
                     /protein_id="AIT38555.1"
                     /translation="METTTTGESADPVTTTVENYGGETQVQRRHHTDVAFVLDRFVEVT
                     VSDNQHTLDVMQAHKDNIVGALLRAATYYFSDLEIAVTHTGKLTWVPNGAPVSALNNTT
                     NPTAYHKGPVTRLALPYTAPHRVLATAYTGTTTYTASARGDSAHLTTTHARHLPTSFNF
                     GAVKAETITELLVRMKRAELYCPRPILPIQPTGDRRKQQLVAPAKQLLGIPSKGEELFT
                     GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGV
                     QCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGNYKTRAEVKFEGDTLVNRIELKG
                     IDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIEDGSVQLADHYQQNT
                     PIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK"
     CDS             909..1619
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      1867..1953
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2045..2072
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2092..2183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2184..3044
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AIT38556.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3215..3803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3989..4129
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        4315..4392
                     /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."
     CDS             4393..5475
                     /codon_start=1
                     /product="lac repressor protein"
                     /label=lac repressor protein
                     /protein_id="AIT38557.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"