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ID PAX4AM preliminary; circular DNA; SYN; 6203 BP.
XX
AC IG1025;
XX
DT 01-FEB-1993 (Rel. 7, Created)
DT 01-JUL-1995 (Rel. 12, Last updated, Version 1)
XX
DE E. coli phagemid vector pAX4a- - complete.
XX
KW cloning vector.
XX
OS Cloning vector
OC Artificial sequences; Cloning vehicles.
XX
RN [1]
RC M13mp11 FX from M13mp11 & oligo, factor X
RC pLmp10 from M13mp10 & pLc245, lambda pL promoter
RC pKG1508 from lambda, nutR/tR1/cII gene
RC pLcII from pLmp10 & pKG1508, lambda nutR/tR1/cII gene
RC M13mp9betaglobin from M13mp9 & pJW102 & human beta-globin
RC pLcIIbeta from pLcII & M13mp9betaglobin & M13mp10
RC plasmid from M13mp11 FX & M13mp9betaglobin
RC pLcIIFXbeta from pLcIIbeta & plasmid
RA Nagai K., Thogersen H.K.;
RT "Generation of beta-globin by sequence-specific proteolysis of a
RT hybrid protein produced in Escherichia coli";
RL Nature 309:810-812(1984).
XX
RN [2]
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
RN [3]
RC pJW101, pJW102, pJW103, pJW112 from pMB9 & pbetaG1, beta-globin gene
RC pJW151, pJW152 from pMB9 & pbetaG1, beta-globin gene
RC from pCR1 & pbetaG1, beta-globin gene
RA Wilson J.T., Wilson L.B., DeRiel J.K., Villa-Komaroff L.,
RA Efstratiadis A., Forget B.G., Weissman S.M.;
RT "Insertion of synthetic copies of human globin genes into
RT bacterial plasmids";
RL Nucleic Acids Res. 5:563-581(1978).
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 (pAX4a-)
CC CM (yes)
CC NA (ds-DNA)
CC TP (circular)
CC ST (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+)(pAX4b+)(pAX4b-)(pAX4c+)(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 39bp
FT \ cgaccatggaattcggtaccagatctagagtcgactgca
FT -> pAX4a- 6203bp"
FT rep_origin complement(0..0)
FT /note="ORI bacteriophage f1 intergenic region"
FT misc_binding 404..404
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 misc_binding 1288..1288
FT /note="SIT unique FspI"
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 6022..6165
FT /note="MCS unique NruI-NcoI-StyI-EcoRI-KpnI-BglII-
FT XbaI-SalI-PstI-SmaI-PacI-HindIII"
FT terminator 6166..6203
FT /note="TER lambda long terminator t0"
XX
SQ Sequence 6203 BP; 1489 A; 1574 C; 1700 G; 1440 T; 0 other;
ttcttgaaga cgaaagggcc accccggttg ataatcagaa aagccccaaa aacaggaaga
ttgtataagc aaatatttaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
taacgaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtgcctcg
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 acagttgcgt 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 gtcgcgacca tggaattcgg taccagatct agagtcgact
gcagcccggg tgattgattg acgatgatta attaattcag aacgctcggt tgccgccggg
cgttttttat gcagcaatgg caagaacgtt gcccggatcc gtcgaagctt atcgatgata
agctgtcaaa catgaagaat taa
//