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ID PAX4CP preliminary; circular DNA; SYN; 6201 BP.
XX
AC IG1030;
XX
DT 01-FEB-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli phagemid vector pAX4c+ - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RC pAX1 from pSS20*c & oligo
RC pAX2 from pAX1 & pSS9
RC pAX3+, pAX3- from pAX2 & pEMBL8+
RC pAX4+ from pAX3+
RC pAX4- from pAX3+
RC pAX4a+ from pAX4+ & linker
RC pAX4b+ from pAX4+ & linker
RC pAX4c+ from pAX4+ & linker
RC pAX4a- from pAX4- & linker
RC pAX4b- from pAX4- & linker
RC pAX4c- from pAX4- & linker
RC pAR10 from pAX1 & E.coli HisRS
RC pPM3220 from pAX4+ & ILS1 gene
RA Markmeyer P., Ruhlmann A., Englisch U., Cramer F.;
RT "The pAX plasmids: new gene-fusion vectors for sequencing,
RT mutagenesis and expression of proteins in Escherichia coli";
RL Gene 93:129-134(1990).
XX
CC ATPG (para-aminophenyl-1-thio-beta-D-galactopyranoside) is like ITPG.
CC Cleave away beta-galactosidase with collagenase or endoproteinase Xa.
CC + makes non-coding strand. - makes coding strand.
CC NM (pAX4c+)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST (non-coding)
CC TY (phagemid)
CC SP (USB)
CC HO (E.coli)
CC CP ()
CC FN (expression)
CC SE (affinity ATPG)
CC PA (pBR322)(f1)
CC BR (pAX4a-)(pAX4a+)(pAX4b-)(pAX4b+)(pAX4c-)(pAX5+)(pAX5-)
CC OF ()
CC OR ()
XX
FH Key Location/Qualifiers
FH
FT misc_feature 0..0
FT /note="1. pBR322 4361bp
FT 2. f1 1174bp
FT -> pSS20*c 5535bp
FT 1. pSS20*c PstI 5535bp
FT 2. oligo IEGR-Xa site-NruI-SalI 30bp
FT \ atcgagggtagggtcgcgatgtcgactgca
FT -> pAX1 5564bp
FT 1. pAX1 remove HindIII-PstI, MCS/5573bp
FT 2. pSS9 NruI-HindIII 84bp, stop codons
FT -> pAX2 5657bp
FT 1. pAX2 DraII 5657bp, MCS
FT alkaline phosphatase
FT 2. pEMBL8+ EcoRII-EcoRI 517bp 3651..3939..229,
FT \ MCS [514bp]
FT -> pAX3+ 6174bp
FT 1. pAX3+ EcoRI 6174bp 0..0
FT mutagenesis
FT -> pAX3+-E5761 6174bp [no EcoRI in lacZ]
FT 1. pAX3+-E5761 EcoRI 6174bp 0..0
FT Klenow
FT -> pAX4+ 6174bp [no EcoRI]
FT 1. pAX4+ remove NruI-PstI 10bp, MCS/6164bp
FT 2. linker NruI-PstI 37bp
FT \ accatggaattcggtaccagatctagagtcgactgca
FT -> pAX4c+ 6201bp"
FT rep_origin 0..0
FT /note="ORI bacteriophage f1 intergenic region"
FT misc_binding 145..145
FT /note="SIT unique NaeI"
FT misc_binding 1030..1030
FT /note="SIT unique ScaI"
FT CDS complement(0..0)
FT /note="ANT E. coli beta-lactamase gene (bla)
FT ampicillin resistance gene (apr/amp)"
FT rep_origin 0..0
FT /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT promoter 0..0
FT /note="PRO E. coli tac (trp and lac)"
FT misc_binding 2979..2979
FT /note="SIT unique Eco81I"
FT CDS 0..0
FT /note="GEN E. coli beta-galactosidase gene (lacZ)
FT alpha peptide"
FT misc_binding 3867..3867
FT /note="SIT unique EcoRV"
FT misc_binding 4083..4083
FT /note="SIT unique BsaBI"
FT misc_binding 4102..4102
FT /note="SIT unique BclI"
FT misc_binding 4254..4254
FT /note="SIT unique BssHII"
FT misc_binding 4589..4589
FT /note="SIT unique Eco47I"
FT misc_binding 4692..4692
FT /note="SIT unique SacI"
FT misc_feature 0..0
FT /note="collagen fragment (CS)"
FT misc_binding 5712..5712
FT /note="SIT unique NdeI"
FT misc_binding 5768..5768
FT /note="SIT unique EspI"
FT misc_binding 5890..5890
FT /note="SIT unique PmlI"
FT misc_binding 5984..5984
FT /note="SIT unique MscI"
FT misc_binding >5984..6021
FT /note="SIT endoproteinase Xa"
FT misc_binding 6026..6163
FT /note="MCS unique NcoI-StyI-EcoRI-KpnI-BglII-
FT XbaI-PstI-SmaI-PacI-HindIII"
FT terminator 6164..6201
FT /note="TER lambda long terminator t0"
XX
SQ Sequence 6201 BP; 1435 A; 1591 C; 1682 G; 1493 T; 0 other;
ttcttgaaga cgaaagggca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
tccacgttcg ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaata
tttgcttata caatcttcct gtttttgggg cttttctgat tatcaaccgg ggtggcctcg
tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
cggaagagcg ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
ttacctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt
cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg tttccggcac
cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg
tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac gtaacctatc
ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca
catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg
ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt cggttacggc caggacagtc
gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc cggagaaaac cgcctcgcgg
tgatggtgct gcgttggagt gacggcagtt atctggaaga tcaggatatg tggcggatga
gcggcatttt ccgtgacgtc tcgttgctgc ataaaccgac tacacaaatc agcgatttcc
atgttgccac tcgctttaat gatgatttca gccgcgctgt actggaggct gaagttcaga
tgtgcggcga gttgcgtgac tacctacggg taacagtttc tttatggcag ggtgaaacgc
aggtcgccag cggcaccgcg cctttcggcg gtgaaattat cgatgagcgt ggtggttatg
ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa actgtggagc gccgaaatcc
cgaatctcta tcgtgcggtg gttgaactgc acaccgccga cggcacgctg attgaagcag
aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa tggtctgctg ctgctgaacg
gcaagccgtt gctgattcga ggcgttaacc gtcacgagca tcatcctctg catggtcagg
tcatggatga gcagacgatg gtgcaggata tcctgctgat gaagcagaac aactttaacg
ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc gaccgctacg
gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca atgaatcgtc
tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga atggtgcagc
gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatgaatca ggccacggcg
ctaatcacga cgcgctgtat cgctggatca aatctgtcga tccttcccgc ccggtgcagt
atgaaggcgg cggagccgac accacggcca ccgatattat ttgcccgatg tacgcgcgcg
tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa tggctttcgc
tacctggaga gacgcgcccg ctgatccttt gcgaatacgc ccacgcgatg ggtaacagtc
ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta cagggcggct
tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc aacccgtggt
cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt atgaacggtc
tggtctttgc cgaccgcacg ccgcatccag cgctgacgga agcaaaacac cagcagcagt
ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac ctgttccgtc
atagcgataa cgagctcctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg
gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac
cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga
ccgcatggtc agaagccggg cacatcagcg cctggcagca gtggcgtctg gcggaaaacc
tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca tctgaccacc agcgaaatgg
atttttgcat cgagctgggt aataagcgtt ggcaatttaa ccgccagtca ggctttcttt
cacagatgtg gattggcgat aaaaaacaac tgctgacgcc gctgcgcgat cagttcaccc
gtgcaccgct ggataacgac attggcgtaa gtgaagcgac ccgcattgac cctaacgcct
gggtcgaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg ttgcagtgca
cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg cagcatcagg
ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt caaatggcga
ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt ggcctgaact
gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg caagaaaact
atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg tcagacatgt
ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc gaattgaatt
atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac agtcaacagc
aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca tggctgaata
tcgacggttt ccatatgggg attggtggcg acgactcctg gagcccgtca gtatcggcgg
tattccagct gagcgccggt cgctaccatt accagttggt ctggtgtcaa aaaggggatc
ctggtcctgt tggtcctgtt ggtcctgctg gtgcttttgg cccaagaggt ctcgctggcc
cacaaggtcc acgtggtgag aaaggtgaac ctggtgataa gggacataga ggtctgcctg
gcctgaaggg acacaatgga ttgcagggtc ttcctggtct tgctggccaa catggtgatc
cgcctgcaat cgagggtagg gtcgaccatg gaattcggta ccagatctag agtcgactgc
agcccgggtg attgattgac gatgattaat taattcagaa cgctcggttg ccgccgggcg
ttttttatgc agcaatggca agaacgttgc ccggatccgt cgaagcttat cgatgataag
ctgtcaaaca tgaagaatta a
//