SBa_000559 vector (V001599)

Basic Vector Information

      • Vector Name:
      • SBa_000559
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5208 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Temme K, Zhao D, Voigt CA.

SBa_000559 vector Vector Map

SBa_0005595208 bp6001200180024003000360042004800SBa_000488SBa_000558SBa_000493SBa_000494cISBa_000496SBa_000497SBa_000498AraCSBa_000500TetRSBa_000491SBa_000502SBa_000503

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

SBa_000559 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5208 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector SBa_000559, complete sequence.
ACCESSION   JQ903615
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5208)
  AUTHORS   Temme K, Zhao D, Voigt CA.
  TITLE     Refactoring the nitrogen fixation gene cluster from Klebsiella 
            oxytoca
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 109 (18), 7085-7090 (2012)
  PUBMED    22509035
REFERENCE   2  (bases 1 to 5208)
  AUTHORS   Temme K, Zhao D, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-APR-2012) Biological Engineering, MIT, 500 Technology 
            Square, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 5208)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5208
                     /organism="Cloning vector SBa_000559"
                     /mol_type="other DNA"
                     /clone="N573"
                     /db_xref="taxon:1176379"
     regulatory      1..42
                     /regulatory_class="terminator"
                     /note="SBa_000488"
     regulatory      43..376
                     /regulatory_class="promoter"
                     /note="SBa_000558"
     misc_feature    43..64
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        65..348
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      377..438
                     /regulatory_class="promoter"
                     /note="SBa_000493"
     protein_bind    377..395
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      439..455
                     /regulatory_class="ribosome_binding_site"
                     /note="SBa_000494"
     CDS             456..1202
                     /codon_start=1
                     /gene="cI"
                     /product="cI"
                     /label=cI
                     /protein_id="AFI71016.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFGAANDENYALVA"
     gene            456..1202
                     /gene="cI"
                     /label=cI
     CDS             456..1166
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=-lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             1167..1199
                     /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"
     regulatory      1239..1373
                     /regulatory_class="terminator"
                     /note="SBa_000496"
     terminator      1247..1318
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1334..1361
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      1374..1416
                     /regulatory_class="promoter"
                     /note="SBa_000497"
     regulatory      1417..1434
                     /regulatory_class="ribosome_binding_site"
                     /note="SBa_000498"
     RBS             1417..1428
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1435..2313
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /protein_id="AFI71017.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            1435..2313
                     /gene="araC"
                     /label=araC
     regulatory      2321..2339
                     /regulatory_class="ribosome_binding_site"
                     /note="SBa_000500"
     CDS             2340..2963
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /protein_id="AFI71018.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            2340..2963
                     /gene="tetR"
                     /label=tetR
     regulatory      2967..3275
                     /regulatory_class="terminator"
                     /note="SBa_000491"
     misc_feature    2967..2987
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     rep_origin      3276..4083
                     /label=SBa_000502
                     /note="SBa_000502"
     rep_origin      complement(3294..3881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3969..4063
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    4084..5208
                     /label=SBa_000503
                     /note="SBa_000503"
     CDS             complement(4084..4944)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4945..5016)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      5205..40
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"

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