LOCUS       Exported                4295 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRS52, complete sequence.
ACCESSION   MG323875
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4295)
  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 4295)
  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 4295)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4295
                     /organism="Cloning vector pRS52"
                     /mol_type="other DNA"
                     /db_xref="taxon:2219087"
     terminator      1..58
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     misc_feature    7..70
                     /label=E. coli His Terminator
                     /note="E. coli His Terminator"
     misc_feature    160..2060
                     /label=rep_ori pSC101 Fragment
                     /note="rep_ori pSC101 Fragment"
     CDS             complement(361..1311)
                     /codon_start=1
                     /product="Rep"
                     /label=Rep
                     /protein_id="AWU50967.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     repeat_region   1313..1359
     regulatory      complement(1342..1347)
                     /regulatory_class="minus_10_signal"
     rep_origin      1359..1581
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     repeat_region   1359..1385
     regulatory      complement(1364..1369)
                     /regulatory_class="minus_35_signal"
     misc_feature    1380..1381
                     /label=ta>at removes SpeI
                     /note="ta>at removes SpeI"
     repeat_region   1409..1428
     repeat_region   1441..1461
     repeat_region   1462..1482
     misc_feature    1480..1562
                     /label=AT Rich Region
                     /note="AT Rich Region"
     misc_feature    complement(1693..2060)
                     /label=par region
                     /note="par region"
     repeat_region   1959..1976
     repeat_region   1982..2005
     repeat_region   2016..2032
     misc_feature    complement(2035..2039)
                     /label=Tn3 integration site
                     /note="Tn3 integration site"
     repeat_region   2045..2058
     regulatory      complement(2066..2171)
                     /regulatory_class="terminator"
                     /note="T0"
     misc_feature    complement(2070..2071)
                     /label=GC>CA
                     /note="GC>CA"
     terminator      2073..2167
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    2197..3245
                     /label=pUC19 fragment
                     /note="pUC19 fragment"
     CDS             complement(2197..3057)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(2197..2859)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AWU50969.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     promoter        complement(3058..3162)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3208..3245
                     /label=BW25113 LacZ Upstream
                     /note="BW25113 LacZ Upstream"
     regulatory      complement(3249..3292)
                     /regulatory_class="terminator"
                     /note="trpT tRNA Terminator"
     terminator      3249..3292
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     regulatory      3295..3440
                     /regulatory_class="promoter"
                     /note="PompF"
     protein_bind    3306..3325
                     /label=OmpR-F1 binding site
                     /bound_moiety="OmpR-F1"
     regulatory      3308..3313
                     /regulatory_class="minus_10_signal"
     protein_bind    3326..3345
                     /label=OmpR-F2 binding site
                     /bound_moiety="OmpR-F2"
     regulatory      3334..3339
                     /regulatory_class="minus_35_signal"
     protein_bind    3346..3365
                     /label=OmpR-F3 binding site
                     /bound_moiety="OmpR-F3"
     regulatory      3354..3359
                     /regulatory_class="minus_35_signal"
     regulatory      3357..3362
                     /regulatory_class="minus_10_signal"
     regulatory      3371..3376
                     /regulatory_class="minus_35_signal"
     regulatory      3382..3387
                     /regulatory_class="minus_10_signal"
     regulatory      3394..3399
                     /regulatory_class="minus_10_signal"
     regulatory      3406
                     /regulatory_class="promoter"
                     /note="+1 nucleotide"
     CDS             3441..4157
                     /codon_start=1
                     /product="superfolder GFP"
                     /label=superfolder GFP
                     /protein_id="AWU50968.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3441..4154
                     /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      4167..4295
                     /regulatory_class="terminator"
                     /note="B0015"
     regulatory      4167..4246
                     /regulatory_class="terminator"
                     /note="B0010"
     terminator      4175..4246
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    4247..4254
                     /label=BB Scar
                     /note="BB Scar"
     regulatory      4255..4295
                     /regulatory_class="terminator"
                     /note="B0012"
     terminator      4262..4289
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"