pJBEI-6409 vector (V007516)

Basic Vector Information

Vector Name:
pJBEI-6409
Antibiotic Resistance:
Chloramphenicol
Length:
14932 bp
Type:
Bacterial Expression, Synthetic Biology ; BglBrick
Replication origin:
p15A ori
Copy Number:
High Copy
Promoter:
PlacUV5
Cloning Method:
Restriction Enzyme
5' Primer:
F5
3' Primer:
R1

pJBEI-6409 vector Map

pJBEI-640914932 bp700140021002800350042004900560063007000770084009100980010500112001190012600133001400014700CAP binding sitelacIlacI promoterpBRrevBamlac UV5 promoterlac operatorAcetyl-CoA acetyltransferaserrnB T1 terminatorT7Te terminatortrc promoterlac operatorMevalonate kinasePhosphomevalonate kinaseDiphosphomevalonate decarboxylaseIsopentenyl-diphosphate Delta-isomerasetrc promoterlac operatorrrnB T1 terminatorT7Te terminatorL4440p15A orilambda t0 terminatorCmRcat promoter

pJBEI-6409 vector Sequence

LOCUS       V007516                14932 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V007516
VERSION     V007516
KEYWORDS    pJBEI-6409
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14932)
  AUTHORS   Alonso-Gutierrez J, Chan R, Batth TS, Adams PD, Keasling JD, Petzold
            CJ, Lee TS
  TITLE     Metabolic engineering of Escherichia coli for limonene and perillyl
            alcohol production.
  JOURNAL   Metab Eng. 2013 May 29;19C:33-41. doi: 10.1016/j.ymben.2013.05.004.
   PUBMED   23727191
REFERENCE   2  (bases 1 to 14932)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 14932)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab
            Eng."; date: "2013-05-29"; pages: "
            10.1016/j.ymben.2013.05.004"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(15..36)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(52..1131)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(1132..1209)
                     /label="lacI promoter"
     primer_bind     1415..1434
                     /label="pBRrevBam"
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
     promoter        1520..1550
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    1558..1574
                     /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             1610..2791
                     /gene="atoB"
                     /label="Acetyl-CoA acetyltransferase"
                     /note="Acetyl-CoA acetyltransferase from Escherichia coli
                     (strain K12). Accession#: P76461"
     terminator      5324..5395
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5411..5438
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     promoter        5650..5679
                     /label="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    5687..5703
                     /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             5729..7057
                     /gene="ERG12"
                     /label="Mevalonate kinase"
                     /note="Mevalonate kinase from Saccharomyces cerevisiae
                     (strain ATCC 204508 / S288c). Accession#: P07277"
     CDS             7081..8433
                     /gene="ERG8"
                     /label="Phosphomevalonate kinase"
                     /note="Phosphomevalonate kinase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P24521"
     CDS             8457..9644
                     /gene="MVD1"
                     /label="Diphosphomevalonate decarboxylase"
                     /note="Diphosphomevalonate decarboxylase from Saccharomyces
                     cerevisiae (strain ATCC 204508 / S288c). Accession#:
                     P32377"
     CDS             9668..10213
                     /gene="idi"
                     /label="Isopentenyl-diphosphate Delta-isomerase"
                     /note="Isopentenyl-diphosphate Delta-isomerase from
                     Escherichia coli (strain K12 / DH10B). Accession#: B1XEH6"
     promoter        10432..10461
                     /label="trc promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    10469..10485
                     /label="lac operator"
                     /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)."
     terminator      13097..13168
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      13184..13211
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(13239..13256)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13373..13918)
                     /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."
     terminator      complement(14032..14126)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(14150..14806)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(14807..14909)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"

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