SBa_000560 vector (V001598)

Basic Vector Information

      • Vector Name:
      • SBa_000560
      • Antibiotic Resistance:
      • Spectinomycin/Streptomycin
      • Length:
      • 9859 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Temme K, Zhao D, Voigt CA.

SBa_000560 vector Vector Map

SBa_0005609859 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600SBa_000491SBa_000492BioBrick prefixlacISBa_000506SBa_000508SBa_000507SBa_000509SBa_000487SBa_000488SBa_000510SBa_000511

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_000560 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9859 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector SBa_000560, complete sequence.
ACCESSION   JQ903616
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9859)
  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 9859)
  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 9859)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9859
                     /organism="Cloning vector SBa_000560"
                     /mol_type="other DNA"
                     /clone="N639"
                     /db_xref="taxon:1176380"
     regulatory      1..92
                     /regulatory_class="terminator"
                     /note="SBa_000491"
     regulatory      93..195
                     /regulatory_class="terminator"
                     /note="SBa_000492"
     terminator      120..149
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    196..217
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(252..1334)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             complement(252..1211)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="AFI71019.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
     gene            complement(252..1211)
                     /gene="lacI"
                     /label=lacI
     regulatory      complement(1212..1909)
                     /regulatory_class="promoter"
                     /note="SBa_000505"
     promoter        complement(1335..1412)
                     /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."
     regulatory      1910..1988
                     /regulatory_class="promoter"
                     /note="SBa_000506"
     protein_bind    1963..1979
                     /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)."
     misc_feature    1989..2028
                     /label=SBa_000508
                     /note="SBa_000508"
     regulatory      2029..2062
                     /regulatory_class="ribosome_binding_site"
                     /note="SBa_000507"
     regulatory      2063..2188
                     /regulatory_class="other"
                     /note="SBa_000509"
     CDS             2180..4837
                     /codon_start=1
                     /product="T7* RNAP"
                     /label=T7* RNAP
                     /protein_id="AFI71020.1"
                     /translation="MGGNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHES
                     YEMGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTA
                     FQFLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKH
                     FKKNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEML
                     IESTGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWT
                     GITGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVA
                     NVITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRIS
                     LEFMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEG
                     YYWLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFC
                     FEYAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIV
                     AKKVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRS
                     VMTLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAA
                     VEAMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMF
                     LGQFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALI
                     HDSFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGN
                     LNLRDILESDFAFA"
     regulatory      4838..4972
                     /regulatory_class="terminator"
                     /note="SBa_000487"
     misc_feature    4838..4858
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4894..4925
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     regulatory      4973..5037
                     /regulatory_class="terminator"
                     /note="SBa_000488"
     rep_origin      5038..8684
                     /label=SBa_000510
                     /note="SBa_000510"
     CDS             5787..6758
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein of plasmid pSa"
                     /label=RepA
                     /translation="MPKNNKAPGHRINEIIKTSLALEMEDAREAGLVGYMARCLVQATM
                     PHTDPKTSYFERTNGIVTLSIMGKPSIGLPYGSMPRTLLAWICTEAVRTKDPVLNLGRS
                     QSEFLQRLGMHTDGRYTATLRNQAQRLFSSMISLAGEQGNDFGIENVVIAKRAFLFWNP
                     KRPEDRALWDSTLTLTGDFFEEVTRSPVPIRIDYLHALRQSPLAMDIYTWLTYRVFLLR
                     AKGRPFVQIPWVALQAQFGSSYGSRARNSPELDDKARERAERAALASFKYNFKKRLREV
                     LIVYPEASDCIEDDGECLRIKSTRLHVTRAPGKGARIGPPPT"
     rep_origin      6795..7230
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    8685..9859
                     /label=SBa_000511
                     /note="SBa_000511"
     CDS             8911..9702
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"

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