Return to this vector's summary.
ID PATH23 preliminary; circular DNA; SYN; 3793 BP.
XX
AC ATCC37703;
XX
DT 01-JUL-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli plasmid vector pATH23 - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RC pATH1 from pKRS101 & linker
RC pATH2 from pATH1 & linker
RC pATH3 from pATH1 & linker
RC pATH10 from pATH1 & pUC8
RC pATH11 from pATH1 & pUC8
RC pATH20 from pATH1 & pUC18
RC pATH21 from pATH1 & pUC18
RC pATH22 from pATH1 & pUC18
RC pATH23 from pATH1 & pUC18
RA Koerner T.J., Hill J.E., Myers A.M., Tzagoloff A.;
RT "High-expression vectors with multiple cloning sites for construction
RT of trpE-fusion genes: pATH vectors";
RL Meth. Enzymol. 194:477-490(1991).
XX
CC Created by Moore, July 1995, under contract with NCBI.
CC The nucleotides remaining in the codon after cutting the vector with
CC the given enzyme are:BamHI-1, ClaI-?, EcoRI-1, HindIII-1, KpnI-2,
CC PstI-2, SacI-2, SalI-1, SmaI-2, SphI-2, XbaI-1, XmaI-3.
CC Restriction digests of the clone give the following sizes (kb):
CC BamHI--4.0; EcoRI--4.0; HindIII--4.0. (ATCC staff)
CC Nucleotide 1 is the first in the trp fragment, at the beginning of a
CC PvuII site 261 nucleotides upstream of the transcription start site.
CC The multiple cloning site follows codon 323 of trpE.
CC To prevent recircularization of the vector, the depositors recommend
CC including 30 - 50 U of calf alkaline phosphatase in the restriction
CC enzyme digestion used to prepare the vector.
CC E.coli containing pATH vectors should be grown on medium
CC supplemented with tryptophan to prevent expression. Storage as DNA at
CC -20C is preferred; long term storage as cells may result in selection
CC of non-expressing promoter mutations.
CC Fusion proteins are produced in E.coli usually in an insoluble
CC form which facilitates purification.Hybrid proteins larger than 90 kDa
CC are not expressed as efficiently as shorter ones.
CC This is one of a series of vectors (ATCC 37695-37703) designed to
CC facilitate expression of an open reading frame as a trpE fusion
CC protein.
CC Medium is -1 1065 plus ampicillin (50 ug/ml) & tryptophan (10 ug/ml).
CC NM (pATH23)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST ()
CC TY (plasmid)
CC SP (ATCC)
CC HO (E.coli RR1)(E.coli)
CC CP ()
CC FN (cloning)
CC SE ()
CC PA (pATH1)(pUC18)(pUC8)
CC BR ()
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pATH1 remove AvaII-AvaII 222bp 2120..2342,
FT \ amp gene 3557bp
FT 2. pUC8 AvaII-AvaII 222bp 1288..1510, amp gene
FT -> plasmid 3779bp [no PstI in amp]
FT 1. plasmid remove SmaI-ClaI 37bp 1421..1458,
FT \ MCS 3742bp
FT blunt end:blunt end
FT 2. pUC18 HindIII-EcoRI 51bp 400..451, MCS
FT blunt end:blunt end
FT -> pATH23 3793bp [no PstI in amp]"
FT - 1..1420
FT /note="pATH1 1..1420 1420bp
FT SmaI = CCC^GGG
FT HindIII = A^AGCTT"
FT - 1421..1471
FT /note="pUC18 400..450 51bp
FT EcoRI = G^AATTC
FT ClaI = AT^CGAT"
FT - 1472..2133
FT /note="pATH1 1458..2119 662bp
FT AvaII = G^GWCC"
FT - 2134..2355
FT /note="pUC8 1288..1509 222bp
FT AvaII = G^GWCC"
FT - 2356..3793
FT /note="pATH1 2342..3779 1438bp"
FT misc_binding 0..0
FT /note="MCS HindIII-SphI-PstI-SalI-XbaI-BamHI-SmaI-
FT KpnI-SacI-EcoRI"
FT misc_binding 0..0
FT /note="SIT unique BamHI-EcoRI-HindIII-KpnI-PstI-
FT SacI-SalI-SmaI-SphI-XbaI-XmaI"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT promoter 0..0
FT /note="PRO E. coli trp gene"
FT CDS 0..0
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amp)"
XX
SQ Sequence 3793 BP; 929 A; 952 C; 958 G; 954 T; 0 other;
cagctgtggt gtcatggtcg gtgatcgcta gggtgccgag cgcatctcga ctgcacggtg
caccaatgct tctggcgtca ggtagttatt ggaaagctgt ggtatggctg tgcaggtcgt
aaatcactgc ataactcgct gctgcctaag gcgcactccc gttctggata atgttttttg
cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
gaactagtta actagtacgc aagttcacgt aaaaagggta tcgacaatga aagcaatttt
cgtactgaaa ggttggtggc gcacttcctg aaacgggcag tgtattcacc atgcgtaaag
caatcagata cccagcccgc ctaatgagcg ggcttttttt tgaacaaaat tagagaataa
caatgcaaac acaaaaaccg actctcgaac tgctaacctg cgaaggcgct tatcgcgaca
atcccaccgc gctttttcac cagttgtgtg gggatcgtcc ggcaacgctg ctgctggaat
ccgcagatat cgacagcaaa gatgatttaa aaagcctgct gctggtagac agtgcgctgc
gcattacagc tttaggtgac actgtcacaa tccaggcact ttccggcaac ggcgaagccc
tcctggcact actggataac gccctgcctg cgggtgtgga aagtgaacaa tcaccaaact
gccgtgtgct gcgcttcccc cctgtcagtc cactgctgga tgaagacgcc cgcttatgct
ccctttcggt ttttgacgct ttccgtttat tgcagaatct gttgaatgta ccgaaggaag
aacgagaagc catgttcttc agcggcctgt tctcttatga ccttgtggcg ggatttgaag
atttaccgca actgtcagcg gaaaataact gccctgattt ctgtttttat ctcgctgaaa
cgctgatggt gattgaccat cagaaaaaaa gcacccgtat tcaggccagc ctgtttgctc
cgaatgaaga agaaaaacaa cgtctcactg ctcgcctgaa cgaactacgt cagcaactga
ccgaagccgc gccgccgctg ccagtggttt ccgtgccgca tatgcgttgt gaatgtaatc
agagcgatga agagttcggt ggcgtagtgc gtttgttgca aaaagcgatt cgcgctggag
aaattttcca ggtggtgcca tctcgccgtt tctctctgcc ctgcccgtca ccgctggcgg
cctattacgt gctgaaaaag agtaatccca gcccgtacat gttttttatg caggataatg
atttcaccct atttggcgcg tcgccggaaa gctcgctcaa gtatgatgcc accagccgcc
agattgagat ccccgggcga gctcgaattc gagctcgccc agcttgcatg cctgcaggtc
gactctagag gatccccggg taccgagctc gcgatgataa gctgtcaaac atgagaatta
attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
gaaacgcgcg agg
//