LOCUS       Exported                5531 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBL119-2, complete sequence.
ACCESSION   MG323871
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5531)
  AUTHORS   Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ.
  TITLE     Phosphatase activity tunes two-component system sensor detection 
            threshold
  JOURNAL   Nat Commun 9 (1), 1433 (2018)
  PUBMED    29650958
REFERENCE   2  (bases 1 to 5531)
  AUTHORS   Landry BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-OCT-2017) Bioengineering, Rice University, 6100 Main 
            St, Houston, TX 77005, USA
REFERENCE   3  (bases 1 to 5531)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5531
                     /organism="Cloning vector pBL119-2"
                     /mol_type="other DNA"
                     /db_xref="taxon:2219082"
     misc_feature    complement(1..6)
                     /label=AvrII Scar
                     /note="AvrII Scar"
     misc_feature    complement(6..819)
                     /label=ColE1 Fragment
                     /note="ColE1 Fragment"
     rep_origin      complement(133..815)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      complement(151..739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(151..703)
                     /label=RNA II
                     /note="RNA II"
     regulatory      557..562
                     /regulatory_class="minus_35_signal"
     regulatory      580..586
                     /regulatory_class="minus_10_signal"
     misc_feature    593..700
                     /label=RNAI
                     /note="RNAI"
     regulatory      complement(711..717)
                     /regulatory_class="minus_10_signal"
     regulatory      complement(735..740)
                     /regulatory_class="minus_35_signal"
     misc_feature    complement(763..815)
                     /label=Tn3 Artifact
                     /note="Tn3 Artifact"
     misc_feature    complement(815..820)
                     /label=SpeI Scar
                     /note="SpeI Scar"
     regulatory      complement(820..925)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      827..921
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    complement(927..1721)
                     /label=CamR Cassette from E. coli R6-5 plasmid
                     /note="CamR Cassette from E. coli R6-5 plasmid"
     misc_feature    complement(927..932)
                     /label=SacI Scar
                     /note="SacI Scar"
     CDS             complement(942..1601)
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /protein_id="AWU50957.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1602..1704)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_feature    1725..1730
                     /label=ClaI
                     /note="ClaI"
     regulatory      1730..1764
                     /regulatory_class="promoter"
                     /note="J23114"
     regulatory      1774..1785
                     /regulatory_class="ribosome_binding_site"
                     /note="B0034"
     RBS             1774..1785
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1793..2503
                     /codon_start=1
                     /product="mCherry"
                     /label=mCherry
                     /protein_id="AWU50955.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    2504..2507
                     /label=Scar
                     /note="Scar"
     regulatory      2508..2559
                     /regulatory_class="terminator"
                     /note="L3S1P52"
     misc_feature    2560..2563
                     /label=Scar
                     /note="Scar"
     misc_feature    2585..2635
                     /label=E. coli LacZ Upstream
                     /note="E. coli LacZ Upstream"
     misc_feature    2637..2642
                     /label=XhoI Scar
                     /note="XhoI Scar"
     promoter        2643..2716
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     regulatory      2643..2696
                     /regulatory_class="promoter"
                     /note="PLTetO-1"
     misc_feature    2643..2661
                     /label=TetR1
                     /note="TetR1"
     protein_bind    2643..2661
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      2662..2667
                     /regulatory_class="minus_35_signal"
     misc_feature    2668..2686
                     /label=TetR2
                     /note="TetR2"
     protein_bind    2668..2686
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      2685..2690
                     /regulatory_class="minus_10_signal"
     misc_feature    2697..2716
                     /label=Lambda Phage Fragment
                     /note="Lambda Phage Fragment"
     misc_feature    2697
                     /label=+1
                     /note="+1"
     regulatory      2717..2747
                     /regulatory_class="ribosome_binding_site"
                     /note="sRBS 3e2"
     CDS             2748..3359
                     /codon_start=1
                     /product="ThsR"
                     /label=ThsR
                     /note="thsR"
                     /protein_id="AWU50953.1"
                     /translation="MQQQINGPVYLVDDDEAIIDSIDFLMEGYGYKLNSFNCGDRFLAE
                     VDLTQAGCVILDARMPGLTGPQVQQLLSDAKSPLAVIFLTGHGDVPMAVDAFKNGAFDF
                     FQKPVPGSLLSQSIAKGLTYSIDQHLKRTNQALIDTLSEREAQIFQLVIAGNTNKQMAN
                     ELCVAIRTIEVHRSKLMTKLGVNNLAELVKLAPLLAHKSE"
     misc_feature    2916..2918
                     /label=Phospho-asp
                     /note="Phospho-asp"
     regulatory      3369..3497
                     /regulatory_class="terminator"
                     /note="B0015"
     regulatory      3369..3448
                     /regulatory_class="terminator"
                     /note="B0010"
     terminator      3377..3448
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    3449..3456
                     /label=BB Scar
                     /note="BB Scar"
     regulatory      3457..3497
                     /regulatory_class="terminator"
                     /note="B0012"
     terminator      3464..3491
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3498..3602
                     /label=Ptrp Fragment
                     /note="Ptrp Fragment"
     regulatory      3615..3806
                     /regulatory_class="promoter"
                     /note="Promoter (Shal_3127)"
     misc_feature    3667..3672
                     /label=half site
                     /note="half site"
     misc_feature    complement(3675..3680)
                     /label=half site
                     /note="half site"
     regulatory      3722..3727
                     /regulatory_class="minus_35_signal"
     regulatory      3742..3747
                     /regulatory_class="minus_10_signal"
     misc_feature    3754
                     /label=+1
                     /note="+1"
     regulatory      3807..3818
                     /regulatory_class="ribosome_binding_site"
                     /note="B0034"
     RBS             3807..3818
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    3819..3824
                     /label=Scar
                     /note="Scar"
     CDS             3825..4541
                     /codon_start=1
                     /product="sgfp"
                     /label=sgfp
                     /protein_id="AWU50956.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3825..4538
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      4557..4685
                     /regulatory_class="terminator"
                     /note="B0015"
     regulatory      4557..4636
                     /regulatory_class="terminator"
                     /note="B0010"
     terminator      4565..4636
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    4637..4644
                     /label=BB Scar
                     /note="BB Scar"
     regulatory      4645..4685
                     /regulatory_class="terminator"
                     /note="B0012"
     terminator      4652..4679
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4686..4730
                     /label=pUC MCS
                     /note="pUC MCS"
     regulatory      4731..4765
                     /regulatory_class="promoter"
                     /note="J23106"
     regulatory      4766..4799
                     /regulatory_class="ribosome_binding_site"
                     /note="sRBS (50,000)"
     CDS             4800..5426
                     /codon_start=1
                     /product="TetR"
                     /label=TetR
                     /note="tetR"
                     /protein_id="AWU50954.1"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     regulatory      5427..5531
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      5432..5518
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"