LOCUS       Exported                5027 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNOR10-20, complete sequence.
ACCESSION   JQ394802
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  AUTHORS   Anderson JC, Voigt CA, Arkin AP.
  TITLE     Environmental signal integration by a modular AND gate
  JOURNAL   Mol. Syst. Biol. 3, 133 (2007)
  PUBMED    17700541
REFERENCE   2  (bases 1 to 5027)
  AUTHORS   Salis HM, Mirsky EA, Voigt CA.
  TITLE     Automated design of synthetic ribosome binding sites to control 
            protein expression
  JOURNAL   Nat. Biotechnol. 27 (10), 946-950 (2009)
  PUBMED    19801975
REFERENCE   3  (bases 1 to 5027)
  AUTHORS   Tamsir A, Tabor JJ, Voigt CA.
  TITLE     Robust multicellular computing using genetically encoded NOR gates 
            and chemical wires
  JOURNAL   Nature 469 (7329), 212-215 (2011)
  PUBMED    21150903
REFERENCE   4  (bases 1 to 5027)
  AUTHORS   Moser F, Voigt CA.
  TITLE     Genetic Circuit Performance under Conditions Relevant for Industrial
            Bioreactors
  JOURNAL   Unpublished
REFERENCE   5  (bases 1 to 5027)
  AUTHORS   Anderson JC, Moser F, Tamsir A, Salis H.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JAN-2012) Biological Engineering, MIT, 500 Tech 
            Square, Rm 209D, Cambridge, MA 02139, USA
REFERENCE   6  (bases 1 to 5027)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5027
                     /organism="Cloning vector pNOR10-20"
                     /mol_type="other DNA"
                     /label=derived from Escherichia coli strain DH10b
                     /note="derived from Escherichia coli strain DH10b"
                     /db_xref="taxon:1175537"
     terminator      78..121
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    124..145
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_feature    124..143
                     /label=biobrick prefix
                     /note="biobrick prefix"
     promoter        146..429
                     /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      146..394
                     /regulatory_class="promoter"
                     /note="PBad"
     protein_bind    458..476
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      506..511
                     /regulatory_class="promoter"
                     /note="Ptet BBa_R0040"
     regulatory      520..530
                     /regulatory_class="ribosome_binding_site"
                     /note="BBa_B0033"
     CDS             537..1283
                     /codon_start=1
                     /product="CI_LVA"
                     /label=CI_LVA
                     /protein_id="AFI26423.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFGAANDENYALVA"
     CDS             537..1247
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=-lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             1248..1280
                     /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      1320..1399
                     /regulatory_class="terminator"
                     /note="BBa_B0015"
     terminator      1328..1399
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1415..1442
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      1455..1489
                     /regulatory_class="promoter"
                     /note="BBa_J23117"
     regulatory      1498..1509
                     /regulatory_class="ribosome_binding_site"
                     /note="BBa_B0034"
     RBS             1498..1509
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1516..2394
                     /codon_start=1
                     /gene="araC"
                     /product="araC"
                     /label=araC
                     /protein_id="AFI26425.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            1516..2394
                     /gene="araC"
                     /label=araC
     CDS             2421..3044
                     /codon_start=1
                     /gene="tetR"
                     /product="tetR"
                     /label=tetR
                     /protein_id="AFI26424.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            2421..3044
                     /gene="tetR"
                     /label=tetR
     misc_feature    3048..3068
                     /label=biobrick suffix
                     /note="biobrick suffix"
     misc_feature    3357..4039
                     /label=colE1
                     /note="colE1"
     rep_origin      complement(3375..3963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4051..4145
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4166..4825)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_feature    complement(4243..4917)
                     /gene="CamR"
                     /label=similar to chloramphenicol acetyltransferase
                     /note="similar to chloramphenicol acetyltransferase"
     gene            complement(4243..4917)
                     /gene="CamR"
                     /label=CamR
     promoter        complement(4826..4928)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"