LOCUS       Exported                9409 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Suicide vector for generating chromosomal deletions in 
            multidrug-resistant bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    pMo130-TelR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9409)
  AUTHORS   Amin IM, Richmond GE, Sen P, Koh TH, Piddock LJ, Chua KL
  TITLE     A method for generating marker-less gene deletions in 
            multidrug-resistant Acinetobacter baumannii.
  JOURNAL   BMC Microbiol. 2013 Jul 13;13:158. doi: 10.1186/1471-2180-13-158.
  PUBMED    23848834
REFERENCE   2  (bases 1 to 9409)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Microbiol."; date: "2013-07-13"; volume: "13:158. doi"; pages: " 
            10.1186/1471-2180-13-158"
FEATURES             Location/Qualifiers
     source          1..9409
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      972..1003
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    complement(1067..1114)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      complement(1293..1881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1373..1392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            2181..2290
                     /note="incP origin of transfer"
     terminator      3541..3586
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    5078..5094
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             complement(5153..5947)
                     /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(5264..5283)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5874..5893
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5948..6051)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6119..6137
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6175..6197)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     promoter        complement(7240..7268)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    complement(7319..7366)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     RBS             7458..7480
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"