LOCUS       Exported               13317 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle cloning vector pDOJ4, complete sequence.
ACCESSION   EF196094
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13317)
  AUTHORS   Lee JH, Halgerson JS, Kim JH, OSullivan DJ.
  TITLE     Comparative sequence analysis of plasmids from Lactobacillus 
            delbrueckii and construction of a shuttle cloning vector
  JOURNAL   Appl. Environ. Microbiol. 73 (14), 4417-4424 (2007)
  PUBMED    17526779
REFERENCE   2  (bases 1 to 13317)
  AUTHORS   Lee J-H., Halgerson JS, Kim J-H., OSullivan DJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-DEC-2006) Center for Microbial and Plant Genomics, 
            University of Minnesota, 1500 Gortner Ave, St. Paul, MN 55108, USA
REFERENCE   3  (bases 1 to 13317)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13317
                     /organism="Shuttle cloning vector pDOJ4"
                     /mol_type="other DNA"
                     /note="Lactobacillus delbrueckii subsp. bulgaricus - 
                     Escherichia coli shuttle vector"
                     /db_xref="taxon:420618"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      647..1493
                     /label=derived from p15A
                     /note="derived from p15A"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1505..1533
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     protein_bind    1986..2007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2022..2052
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2060..2076
                     /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)."
     primer_bind     2084..2100
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2096..2419
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /note="derived from pUC18 for blue/white screening"
                     /protein_id="ABM79766.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     gene            2096..2419
                     /gene="lacZ"
                     /label=lacZ
     misc_feature    2110..2166
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    2112..2168
                     /gene="lacZ"
                     /label=multiple cloning site of LacZ from pUC18
                     /note="multiple cloning site of LacZ from pUC18"
     primer_bind     complement(2170..2186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2660..2764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(4044..4055)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4518..4976
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="derived from pCI372 for chloramphenicol resistance"
                     /protein_id="ABM79767.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPEKCFFSFYYSMDFIYWV"
     gene            4518..4976
                     /gene="cat"
                     /label=cat
     misc_feature    6796..13015
                     /label=pDOJ1 region
                     /note="pDOJ1 region"
     CDS             7073..7651
                     /codon_start=1
                     /product="putative mobilization protein"
                     /label=putative mobilization protein
                     /note="ORF1; derived from Lactobacillus delbrueckii subsp. 
                     bulgaricus plasmid pDOJ1"
                     /protein_id="ABM79768.1"
                     /translation="MENSAGMLSVGQIAKMLKTNRQNIYNVLKTEHIKADGYNDKHYSL
                     YSPETIQEIKAALSKKATLRSKKVVAKEQAEEIADLKNQLSEQQRLTTWLQSQLVQLQV
                     EADKLRSQNSQLQLDHAKTQLLIGQVDQEKTTLKAENDRLSAENDKLGQLTDKVLKDAQ
                     RAEEDAQKAKADLDKARRAGLWSRITRNY"
     CDS             7952..8473
                     /codon_start=1
                     /product="putative transposase"
                     /label=putative transposase
                     /note="ORF2; derived from Lactobacillus delbrueckii subsp. 
                     bulgaricus plasmid pDOJ1"
                     /protein_id="ABM79769.1"
                     /translation="MTYKEWTDSDHLELVKNWKLHGLTNVEIAQRIGIAEKTLYVWLKK
                     SPKLKKAIRGGKNIARARAENALYELALNGDRQALFFWLKNNYRERYSDKPLSPAEADL
                     MSQKARLAKLQADLAEAQLKAIKEDQGDQATQLNNLLDSLKEAVLDEGISPDNIVPTGN
                     GLIIDDISDS"
     CDS             9128..9322
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF3; derived from Lactobacillus delbrueckii subsp. 
                     bulgaricus plasmid pDOJ1"
                     /protein_id="ABM79770.1"
                     /translation="MIRQNSDVRAAIKASGLKQYEVATLMNVSASYLSQLLLQPLSEGH
                     KKRIMAAIKQGESLKGEQE"
     CDS             9301..9312
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             9322..9549
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF4; derived from Lactobacillus delbrueckii subsp. 
                     bulgaricus plasmid pDOJ1"
                     /protein_id="ABM79771.1"
                     /translation="MMSLEEREREIEKVVRIAEADFNNAYHLNAINKEDVIKNRAYKYA
                     EVLKLQELLALNKTIKDGLNGIEMSVDLIE"
     CDS             9564..10535
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF5; derived from Lactobacillus delbrueckii subsp. 
                     bulgaricus plasmid pDOJ1"
                     /protein_id="ABM79772.1"
                     /translation="MNNSEKTSLMAEPYNSNRNAIDRLRVNQKALQAGSVKREENYIAD
                     ESGNSEGFRMAFYTAYKSGVQLAIPVEAEPGKMVINETFSKVQHLLIASWYSQADRASN
                     FRIQLTFKEISEALGYSRSQATALRKQLRDLITPLVRCTFVTSNKDSIDAVNLFAAGSY
                     SKGQLTMWLTPNMAETFLSEESSTEYFPLSLLKLKGTAYYLALKIMHNANINARWRADR
                     VDRLGLEDTLKALPTLPDPVENAQGTSRHLQRQIIAPLAKAIEELEAVTGIVVRPSQPL
                     KGLKAKDLSKVTLNVTDWGQVDIAELTRKKRKRLRKNNVRED"
     rep_origin      10558..10622
                     /label=derived from pDOJ1
                     /note="derived from pDOJ1"
     repeat_region   10558..10578
                     /label=direct repeat I for iteron structure
                     /note="direct repeat I for iteron structure"
     repeat_region   10580..10600
                     /label=direct repeat II for iteron structure
                     /note="direct repeat II for iteron structure"
     misc_feature    10602..10622
                     /label=direct repeat III for iteron structure
                     /note="direct repeat III for iteron structure"
     CDS             10830..12896
                     /codon_start=1
                     /product="putative primase-helicase"
                     /label=putative primase-helicase
                     /note="ORF6; derived from Lactobacillus delbrueckii subsp. 
                     bulgaricus plasmid pDOJ1"
                     /protein_id="ABM79773.1"
                     /translation="MNNLVKPADLKSLISLPEYIASVVSMDSKGFFSCVNPNHPDKHPS
                     MCVDQNHPQYVHCFSCQASYDLFDCWALINDGVTETRKNSAGKEKPVYNFNAVAIEIAD
                     HYGYTLIGDPANNLPEPPLPEPEPEPAQTQTSNDSNIRAQLEAWHASLNQTDYLQKRGI
                     TQTTAEIFNLGYSALTNSIIIPYGQDGYYVQRAINPIEKHDRYRFPIGQVRVYNAEALK
                     ECKTVFIVEGQFDALSIMQESGVGAVATSTSQTRLIVKTLQKFKEQDPTINPTIILSMD
                     NDRAGQKANRALQRDLEAHGFTCYVNPVNGDYKDANEFLVKDREGFRQKLQHVINQPDN
                     WLDKYYADIKNRHDYPDNIPTGFKNLDDELDGGLQPKLYVLGAVSSLGKTTFALNIADN
                     LAKQGRHVFFFSMESSKREVTDKLLSRASCLSNGHKWTQLQVSRGAWLNNAEDKEEFDG
                     LFKAFSRYQRFLHIYDNRVKASQVKDLVNSWLGNHPDEKKPLVVVDYLQILQAEQDNVT
                     DKAKVTDSVSVLSELTKQAEVPVLVISSLNRASYWQDVSFESFKESGEIEYSADVMLGL
                     EFAHREEYITVQKNGHVELNKEKFDQRKQEVPRRVEMVILKNRTGKTGGHIFFKYNAMF
                     NSYQACTEKEAAIANNFKMLYHTKQVGKPVEAARIEYTVDPETGRVTEKNQDK"