LOCUS       Exported                5009 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Boolean integrase logic vector NOR gate BCD-RFP, complete sequence.
ACCESSION   KM234322
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5009)
  AUTHORS   Courbet A, Endy D, Renard E, Molina F, Bonnet J.
  TITLE     Detection of pathological biomarkers in human clinical samples via 
            amplifying genetic switches and logic gates
  JOURNAL   Sci Transl Med 7 (289), 289RA83 (2015)
  PUBMED    26019219
REFERENCE   2  (bases 1 to 5009)
  AUTHORS   Courbet A, Molina F, Endy D, Bonnet J.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUL-2014) SysDiag CNRS UMR3145, CNRS, 1682 Rue de la 
            Valsiere, Montpellier, Herault 34184, France
REFERENCE   3  (bases 1 to 5009)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5009
                     /organism="Boolean integrase logic vector NOR gate
                     BCD-RFP"
                     /mol_type="other DNA"
                     /clone="NOR BCD RFP"
                     /db_xref="taxon:1642391"
     misc_feature    1..22
                     /label=prefix_Bba
                     /note="prefix_Bba"
     regulatory      23..69
                     /regulatory_class="promoter"
                     /note="P7"
     misc_feature    70..122
                     /label=attB TP901-1
                     /note="attB TP901-1"
     regulatory      123..251
                     /regulatory_class="terminator"
                     /note="B0015"
     terminator      129..156
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      172..243
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    252..301
                     /label=attP TP901-1
                     /note="attP TP901-1"
     misc_feature    302..351
                     /label=attB Bxb1
                     /note="attB Bxb1"
     regulatory      352..464
                     /regulatory_class="terminator"
                     /note="J61048"
     misc_feature    465..517
                     /label=attP Bxb1
                     /note="attP Bxb1"
     misc_feature    518..592
                     /label=RiboJ
                     /note="RiboJ"
     misc_RNA        520..569
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_feature    593..677
                     /label=BCD2
                     /note="BCD2"
     RBS             663..671
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             678..1373
                     /codon_start=1
                     /product="mKate2"
                     /label=mKate2
                     /protein_id="AKC96471.1"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     regulatory      1385..1513
                     /regulatory_class="terminator"
                     /note="B0015"
     terminator      1393..1464
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1480..1507
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1514..1534
                     /label=suffix_Bba
                     /note="suffix_Bba"
     terminator      1570..1601
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      1697..3733
                     /label=pSC101
                     /note="pSC101"
     terminator      1726..1820
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2312..2534
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2582..3532
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     misc_feature    3742..4684
                     /label=Amp resistance
                     /note="Amp resistance"
     CDS             3821..4681
                     /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"
     terminator      4811..4840
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      4934..4963
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"