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"