LOCUS EF372397 5534 bp DNA circular SYN 26-APR-2007
DEFINITION Expression vector pOPINE, complete sequence.
ACCESSION EF372397
VERSION EF372397.1
KEYWORDS .
SOURCE Expression vector pOPINE
ORGANISM Expression vector pOPINE
other sequences; artificial sequences; vectors.
REFERENCE 1 (bases 1 to 5534)
AUTHORS Berrow,N.S., Alderton,D., Sainsbury,S., Nettleship,J.,
Assenberg,R., Rahman,N., Stuart,D.I. and Owens,R.J.
TITLE A versatile ligation-independent cloning method suitable for
high-throughput expression screening applications
JOURNAL Nucleic Acids Res. 35 (6), E45 (2007)
PUBMED 17317681
REFERENCE 2 (bases 1 to 5534)
AUTHORS Berrow,N.S.
TITLE Direct Submission
JOURNAL Submitted (17-JAN-2007) Oxford Protein Production Facility, Oxford
University, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, U.K.
FEATURES Location/Qualifiers
source 1..5534
/organism="Expression vector pOPINE"
/mol_type="other DNA"
/db_xref="taxon:425270"
/note="based on pTriEx2 backbone"
misc_feature 45..1011
/note="ORF603/lef2"
regulatory 1079..1443
/regulatory_class="enhancer"
/note="CMV enhancer"
regulatory 1449..1726
/regulatory_class="promoter"
/note="chicken beta-actin promoter"
regulatory 2150..2166
/regulatory_class="promoter"
/note="T7 promoter"
misc_feature 2171..2191
/note="lac operator"
misc_feature 2205..2318
/note="p10 promoter and 5' UTR"
regulatory 2319..2326
/regulatory_class="ribosome_binding_site"
misc_feature 2321..2335
/note="5' in-fusion site (after NcoI digest)"
misc_feature 2369..2699
/note="lacZ promoter and lacZ alpha peptide"
misc_feature 2719..2739
/product="KHHHHHH tag"
misc_feature complement(2719..2733)
/note="3' in-fusion site (after PmeI digest)"
regulatory 2797..3033
/regulatory_class="polyA_signal_sequence"
regulatory 3037..3084
/regulatory_class="terminator"
/note="T7 terminator"
misc_feature 3097..3802
/note="ORF1629"
rep_origin 4013
/note="pUC origin of replication"
gene complement(4612..5472)
/gene="ampR"
CDS complement(4612..5472)
/gene="ampR"
/codon_start=1
/transl_table=11
/product="beta-lactamase"
/protein_id="ABN80182.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW"
BASE COUNT 1404 a 1293 c 1272 g 1565 t
ORIGIN
1 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga
61 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
121 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
181 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
241 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
301 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
361 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
421 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
481 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
541 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
601 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
661 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
721 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
781 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
841 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
901 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
961 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggccg
1021 taatgagacg cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctct
1081 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
1141 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
1201 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
1261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
1321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
1381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
1441 atgcatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
1501 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
1561 gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
1621 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
1681 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
1741 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
1801 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctccttcgg gctgtaatta
1861 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
1921 ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc
1981 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
2041 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
2101 ttattgtgct gtctcatcat tttggcaaag aattggatcg gaccgaaatt aatacgactc
2161 actatagggg aattgtgagc ggataacaat tccccggagt taatccggga cctttaattc
2221 aacccaacac aatatattat agttaaataa gaattattat caaatcattt gtatattaat
2281 taaaatacta tactgtaaat tacattttat ttacaatcaa aggagatata ccatggcaca
2341 ccatcaccac catcacagca gcggtaccgt cgacccgact ggaaagcggg cagtgagcgc
2401 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
2461 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
2521 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
2581 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
2641 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcca
2701 agctttctag accagtttaa acatcaccat caccatcact aagtgattaa cctcaggtgc
2761 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
2821 gagatcgatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
2881 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
2941 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
3001 ttggtttaga gtttggcaac atatgcccat atgtaactag cataacccct tggggcctct
3061 aaacgggtct tgaggggttt tttgctgaaa gcatgcggag gaaattctcc ttgaagtttc
3121 cctggtgttc aaagtaaagg agtttgcacc agacgcacct ctgttcactg gtccggcgta
3181 ttaaaacacg atacattgtt attagtacat ttattaagcg ctagattctg tgcgttgttg
3241 atttacagac aattgttgta cgtattttaa taattcatta aatttataat ctttagggtg
3301 gtatgttaga gcgaaaatca aatgattttc agcgtcttta tatctgaatt taaatattaa
3361 atcctcaata gatttgtaaa ataggtttcg attagtttca aacaagggtt gtttttccga
3421 accgatggct ggactatcta atggattttc gctcaacgcc acaaaacttg ccaaatcttg
3481 tagcagcaat ctagctttgt cgatattcgt ttgtgttttg ttttgtaata aaggttcgac
3541 gtcgttcaaa atattatgcg cttttgtatt tctttcatca ctgtcgttag tgtacaattg
3601 actcgacgta aacacgttaa atagagcttg gacatattta acatcgggcg tgttagcttt
3661 attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta gaagttgctt ccgaagacga
3721 ttttgccata gccacacgac gcctattaat tgtgtcggct aacacgtccg cgatcaaatt
3781 tgtagttgag ctttttggaa ttgcgatcgc ataacttcgt atagcataca ttatacgaag
3841 ttataagctc ggaacgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
3901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
3961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
4021 gctccgcccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
4081 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
4141 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
4201 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
4261 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
4321 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
4381 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
4441 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
4501 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
4561 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gttaccaatg
4621 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
4681 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
4741 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
4801 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
4861 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
4921 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
4981 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
5041 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
5101 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
5161 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
5221 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
5281 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
5341 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
5401 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
5461 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
5521 catgtccgcg cgtt
//