LOCUS       Exported                4946 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  low copy plasmid for expression of a variant of the fluorescent 
            timer protein suitable for bacteria under the control of the 
            constitutive ybaJp promoter (works in E. coli and Salmonella 
            enterica).
ACCESSION   .
VERSION     .
KEYWORDS    pSC101_TIMER
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4946)
  AUTHORS   Claudi B, Sprote P, Chirkova A, Personnic N, Zankl J, Schurmann N, 
            Schmidt A, Bumann D
  TITLE     Phenotypic variation of Salmonella in host tissues delays 
            eradication by antimicrobial chemotherapy.
  JOURNAL   Cell. 2014 Aug 14;158(4):722-733. doi: 10.1016/j.cell.2014.06.045.
  PUBMED    25126781
REFERENCE   2  (bases 1 to 4946)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2014.06"; journalName: "Cell"; date: "2014-08-14- 
            14"; volume: "158"; issue: "4"; pages: "722-733"
FEATURES             Location/Qualifiers
     source          1..4946
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      64..158
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      650..872
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             920..1870
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      2408..2494
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2622..2641
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3416..3434)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3466..3486)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3684..3703
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3878..3901
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3988..4006)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             join(4185..4946,1..33)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4239..4258)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4849..4868
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"