pRP1028-T1-PL vector (V016234)

Basic Vector Information

Vector Name:
pRP1028-T1-PL
Length:
8081 bp
Type:
Cloning vector
Replication origin:
pSC101 ori
Source/Author:
Bernhards CB, Lux MW, Katoski SE, Goralski TDP.

pRP1028-T1-PL vector Map

pRP1028-T1-PL8081 bp400800120016002000240028003200360040004400480052005600600064006800720076008000I-SceI siteoriTT7 promoterM13 fwdts rep from pJRS233, pWV01Factor Xa sitePtrcPtrcregulatorySpectinomycin 9-adenylyltransferasePrrnBPrrnBPFP2PFP2regulatoryTurboRFPRep101regulatorypSC101 oriparBtk_IntergenicTarget1_Polylinker

pRP1028-T1-PL vector Sequence

LOCUS       V016234                 8081 bp    DNA     circular SYN 14-MAR-2021
DEFINITION  Exported.
ACCESSION   V016234
VERSION     V016234
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8081)
  AUTHORS   Bernhards CB, Lux MW, Katoski SE, Goralski TDP., Liem AT, Gibbons
            HS.
  TITLE     barCoder: a tool to generate unique, orthogonal genetic tags for
            qPCR detection
  JOURNAL   BMC Bioinformatics 22 (1), 98 (2021)
   PUBMED   33648451
REFERENCE   2  (bases 1 to 8081)
  AUTHORS   Bernhards CB, Lux MW, Katoski SE, Goralski TDP., Liem AT, Gibbons
            HS.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2020) US Army CCDC Chemical Biological Center,
            8198 Blackhawk Rd, Gunpowder, MD 21010, USA
REFERENCE   3  (bases 1 to 8081)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "BMC
            Bioinformatics"; date: "2021"; volume: "22"; issue: "1"; pages: "98"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-SEP-2020) US Army CCDC Chemical Biological Center, 8198
            Blackhawk Rd, Gunpowder, MD 21010, USA"
FEATURES             Location/Qualifiers
     source          1..8081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    12..29
                     /label="I-SceI site"
                     /note="I-SceI site"
     oriT            180..288
                     /label="oriT"
                     /note="incP origin of transfer"
     promoter        complement(303..321)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(331..347)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      909..2231
                     /label="ts rep from pJRS233, pWV01"
                     /note="ts rep from pJRS233, pWV01"
     gene            complement(909..1058)
                     /gene="orfD"
                     /label="orfD"
     CDS             complement(909..1058)
                     /codon_start=1
                     /transl_table=11
                     /gene="orfD"
                     /product="OrfD"
                     /label="orfD"
                     /protein_id="QSS48602.1"
                     /translation="MTNKEKELFAENEELKKEIKDLKERIERYREMEVELSTTIDLLRG
                     GIIE"
     gene            complement(1055..1753)
                     /gene="repA"
                     /label="repA"
     CDS             complement(1055..1753)
                     /codon_start=1
                     /transl_table=11
                     /gene="repA"
                     /product="RepA"
                     /label="repA"
                     /protein_id="QSS48609.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     misc_difference 1578..1595
                     /note="mutations in repA leading to temperature
                     sensitivity"
     gene            complement(1820..1981)
                     /gene="orfC"
                     /label="orfC"
     CDS             complement(1820..1981)
                     /codon_start=1
                     /transl_table=11
                     /gene="orfC"
                     /product="OrfC"
                     /label="orfC"
                     /protein_id="QSS48603.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     gene            complement(2022..2231)
                     /gene="orfB"
                     /label="orfB"
     CDS             complement(2022..2231)
                     /codon_start=1
                     /transl_table=11
                     /gene="orfB"
                     /product="OrfB"
                     /label="orfB"
                     /protein_id="QSS48604.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     CDS             2365..2376
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     regulatory      2551..2556
                     /label="Ptrc"
                     /note="Ptrc"
                     /regulatory_class="minus_35_signal"
     regulatory      2574..2578
                     /label="Ptrc"
                     /note="Ptrc"
                     /regulatory_class="minus_10_signal"
     regulatory      2594..2603
                     /regulatory_class="ribosome_binding_site"
     CDS             2611..3390
                     /gene="ant1"
                     /label="Spectinomycin 9-adenylyltransferase"
                     /note="Spectinomycin 9-adenylyltransferase from
                     Staphylococcus aureus (strain N315). Accession#: P0A0D1"
     regulatory      3419..3424
                     /label="PrrnB"
                     /note="PrrnB"
                     /regulatory_class="minus_35_signal"
     regulatory      3442..3447
                     /label="PrrnB"
                     /note="PrrnB"
                     /regulatory_class="minus_10_signal"
     regulatory      3467..3472
                     /label="PFP2"
                     /note="PFP2"
                     /regulatory_class="minus_35_signal"
     regulatory      3490..3495
                     /label="PFP2"
                     /note="PFP2"
                     /regulatory_class="minus_10_signal"
     regulatory      3508..3517
                     /regulatory_class="ribosome_binding_site"
     CDS             3544..4236
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     CDS             complement(4767..5714)
                     /label="Rep101"
                     /note="RepA protein needed for replication with the pSC101
                     origin"
     regulatory      complement(5745..5750)
                     /regulatory_class="minus_10_signal"
     rep_origin      complement(5762..5984)
                     /direction=LEFT
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     misc_feature    complement(6095..6463)
                     /label="par"
                     /note="par"
     misc_feature    complement(6478..8073)
                     /label="Btk_IntergenicTarget1_Polylinker"
                     /note="Btk_IntergenicTarget1_Polylinker"
     misc_feature    complement(join(6484..7169,7206..8069))
                     /note="similar to Bacillus thuringiensis serovar kurstaki
                     sequence in accession NZ_CP010005.1"
     misc_feature    6484..6988
                     /note="similar to fatty acid--CoA ligase; BG08_RS29910"
     misc_feature    7170..7205
                     /label="polylinker"
                     /note="polylinker"
     misc_feature    7170..7175
                     /label="XhoI restriction site"
                     /note="XhoI restriction site"
     misc_feature    7176..7181
                     /label="SmaI restriction site"
                     /note="SmaI restriction site"
     misc_feature    7182..7187
                     /label="SphI restriction site"
                     /note="SphI restriction site"
     misc_feature    7188..7193
                     /label="NheI restriction site"
                     /note="NheI restriction site"
     misc_feature    7194..7199
                     /label="NcoI restriction site"
                     /note="NcoI restriction site"
     misc_feature    7200..7205
                     /label="SacI restriction site"
                     /note="SacI restriction site"
     CDS             complement(7573..7770)
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label="hypothetical protein"
                     /note="similar to BG08_RS29905"
                     /protein_id="QSS48608.1"
                     /translation="MNTTAKLINWKEHGDMIILECELNGKRFEISTYKQRIYNAHLLSD
                     DVYIRLDSSDNIIGINIYKK"
     misc_feature    complement(7905..8069)
                     /note="similar to hypothetical protein; BG08_RS29900"

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