DH7-21 vector (V010012)

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LOCUS       40924_650        7582 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DH7-21, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7582)
  AUTHORS   Huang DC, Holtz WJ, Maharbiz MM.
  TITLE     A genetic bistable switch utilizing nonlinear protein degradation
  JOURNAL   J Biol Eng 6 (1), 9 (2012)
  PUBMED    22776405
REFERENCE   2  (bases 1 to 7582)
  AUTHORS   Huang DC, Holtz WJ, Maharbiz MM.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUN-2012) Electrical Engineering and Computer 
            Sciences, University of California, Berkeley, 656 Sutardja Dai Hall,
            Berkeley, CA 94720, USA
REFERENCE   3  (bases 1 to 7582)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7582)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Eng"; date: "2012"; volume: "6"; issue: "1"; pages: "9"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-JUN-2012) Electrical Engineering and Computer Sciences, 
            University of California, Berkeley, 656 Sutardja Dai Hall, Berkeley,
            CA 94720, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: GENtle v. 1.9.4
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            complement(1..2400)
                     /gene="mf-lon with LVA"
                     /label=mf-lon with LVA
     CDS             complement(4..2400)
                     /codon_start=1
                     /gene="mf-lon with LVA"
                     /product="mf Lon with LVA degradation tag"
                     /label=mf-lon with LVA
                     /protein_id="AFN88297.1"
                     /translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
                     QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
                     DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
                     DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
                     QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
                     RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
                     KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
                     DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
                     PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
                     DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
                     EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
                     NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG
                     ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
                     ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTKAANDENYALVA"
     misc_feature    complement(7..39)
                     /gene="mf-lon with LVA"
                     /label=degradation tag (LVA)
                     /note="degradation tag (LVA)"
     CDS             complement(7..39)
                     /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"
     5'UTR           complement(2401..2434)
     protein_bind    2445..2463
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(2470..2488)
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      2495..2576
                     /label=pRM
                     /note="pRM"
                     /regulatory_class="promoter"
     5'UTR           2583..2617
     CDS             2618..3328
                     /label=lambda repressor
                     /note="phage lambda repressor"
     misc_feature    3329..3409
                     /gene="cI-mf-ssra tag"
                     /label=mf-Lon ssrA degradation tag
                     /note="mf-Lon ssrA degradation tag"
     5'UTR           3422..3462
     CDS             3463..4083
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             4084..4116
                     /label=ssrA tag (LVA)
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
     terminator      4155..4226
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4242..4269
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4803..5750)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(5798..6020)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(6512..6606)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6640..7431)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"