pS239D.M vector (V016197)

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LOCUS       62056_19150        8316 bp DNA     circular SYN 26-NOV-2019
DEFINITION  Expression vector pS239D.M, complete sequence.
ACCESSION   MN393464
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  AUTHORS   Calles B, Goni-Moreno A, de Lorenzo V.
  TITLE     Digitalizing heterologous gene expression in Gram-negative bacteria 
            with a portable on/off module
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8316)
  AUTHORS   Calles B, Goni-Moreno A, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2019) Systems Biology Program, Centro Nacional de 
            Biotecnologia, Darwin, 3, Cantoblanco, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 8316)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-AUG-2019) Systems Biology Program, Centro Nacional de 
            Biotecnologia, Darwin, 3, Cantoblanco, Madrid 28049, Spain"
FEATURES             Location/Qualifiers
     source          1..8316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5..99
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             231..1043
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     oriT            1213..1321
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1335..1994)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(1995..2766)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(2881..2967)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     gene            complement(2982..5639)
                     /gene="alkS"
                     /label=alkS
     CDS             complement(2982..5639)
                     /codon_start=1
                     /transl_table=11
                     /gene="alkS"
                     /product="AlkS"
                     /label=alkS
                     /note="edited; crc mut"
                     /protein_id="QGN03500.1"
                     /translation="MGMKIKIINNDFPVAKIGLDRITTLVSAKVHNCIYRPRLSIADGT
                     APRVCLYRAPPGYGKTVALAFEWLRHRTTGRPAVWISLRASSYSEFDICAEIIEQLEAF
                     ELVTFSHVREGVSKPTLLRDLASSLWQSTSSNEIETLICLDNINQGLGLPLLHALMEFM
                     LETPKSIRFAVAGNTIKGFSRLKLAGAMQEHTEKDLAFSADEAVALVEAEAVLGVSEVQ
                     IEALVQEMEGWPVLIGFLLKCELPAKSISTVVEIDNYFNDEIFEALPERYRVFLVNSSL
                     LDVVTPDGYNYVFKCVNAASCIKYLSTNYMLLRHVEGEPAQFTLHPVLRDFLQGIAWAE
                     NPAKRSYLLKRAAFWHWRRGEYQYAIRIALRANDCRWAVGMSEGIILDLSFRQGEIDTL
                     RHWLSELPVKDLHKNPIVLICFAWVLYFSQQSARAEKLLKDLITPPDKKNKWQEKGWPQ
                     LVFAIGKATNDEMLLSEELCNKWISLFGDSNAVGKGAALTCLAFIFASEYRFAELEKVL
                     AQAQAVNKFAKQDFAFGWLYVAKLQQALASGKMSWARQLITQARTDSGAQIMETAFTSK
                     MLDALELESNYELCRLDTSEEKFSEILEFIANHGVTDVFFSVCRVVSAWRLGRNDLNGS
                     IEILEWAKAYAAEKNLPRLEVMSQIEIYQRLLFQGVTYINTLQAFEDRKIFSGPHSAPL
                     KARLLLVQSLALSRDQNFHLAAHRALLAIQQARKISAGQLEVRGLLYLAGAQAGGGTLK
                     KAQHNIAYALEMAKQLQCFQTVLDEIRLIKRLIPTSCEVFAAVNLDQAIGAFSLPQMVE
                     IRKSAENKAGDFLTLKQVSVLKLVKEGCSNKQIATKMYVTEDAIKWHMRKIFTILNVKS
                     RTQAIIEAERQGVI"
     regulatory      complement(5666..5771)
                     /label=Km promoter
                     /note="Km promoter"
                     /regulatory_class="promoter"
     regulatory      5975..6035
                     /label=PAlkB
                     /note="PAlkB"
                     /regulatory_class="promoter"
     regulatory      6039..6045
                     /regulatory_class="ribosome_binding_site"
     CDS             6057..7136
                     /label=lacI
                     /note="lac repressor"
     CDS             7137..7154
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             7183..7893
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     misc_feature    7897..7953
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      7986..8015
                     /label=synthetic T500 terminator
                     /note="synthetic T500 terminator"
                     /regulatory_class="terminator"
     terminator      complement(8022..8049)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(8089..8135)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      complement(8150..8258)
                     /note="anti LacI sRNA; MicC-based"
                     /regulatory_class="other"
     regulatory      complement(8259..8316)
                     /label=PA1/O4s
                     /note="PA1/O4s"
                     /regulatory_class="promoter"