LOCUS       Exported                8859 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Suicide plasmid for efficient gene mutation in Gram-positive 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    pHoss1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  AUTHORS   Abdelhamed H, Lawrence ML, Karsi A
  TITLE     A novel suicide plasmid for efficient gene mutation in Listeria 
            monocytogenes.
  JOURNAL   Plasmid. 2015 May 31;81:1-8. doi: 10.1016/j.plasmid.2015.05.003.
  PUBMED    26038185
REFERENCE   2  (bases 1 to 8859)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid."; 
            date: "2015-05-31"; volume: "81:1-8. doi"; pages: " 
            10.1016/j.plasmid.2015.05.003"
FEATURES             Location/Qualifiers
     source          1..8859
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(4069..4692)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRRHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     4600..4619
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    4717..4735
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    4802..4820
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             6416..6607
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6601..6623)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     misc_feature    6709..6849
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6764..6783
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6864..6881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7035..7623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7115..7134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7794..8654)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8417..8436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8655..8759)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8827..8845
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"