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"