pEE12.4
Search name
pEE12.4,Plasmid pEE12.4,pEE12.4 vector
Promoter: hCMV-MIE
Replicator: Pee6 ori
Terminator: SV40, poly (A) signal
Plasmid classification: mammalian cells, antibody expression vectors
Plasmid size: 7569bp
Prokaryotic resistance: Amp
Clone strain: DH5 alpha
Culture conditions: 37 DEG C, aerobic LB
Expression host: mammalian cells
Induction mode: no induction, transient expression
Primers for 5'sequencing: CMV-F:CGCAAATGGGCGGTAGGCGTG
Primers for 3'sequencing: according to sequences
pEE12.4 Sequence
LOCUS Exported 7569 bp ds-DNA circular SYN 10-AUG-2017
FEATURES Location/Qualifiers
source 1..7569
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 116..237
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(999..1587)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1758..2618)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2619..2723)
/gene="bla"
/label=AmpR promoter
terminator 2892..2919
/label=T7Te terminator
/note="phage T7 early transcription terminator"
promoter 3002..3331
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3182..3317
/label=SV40 ori
/note="SV40 origin of replication"
intron 4623..4688
/label=small t intron
/note="simian virus 40 (SV40) small t antigen intron"
CDS 4818..4838
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5263..5397
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
enhancer 5966..6345
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 6346..6549
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
ORIGIN
1 gaattcattg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
61 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
241 tgtctggcgg ccgcgacctg caggcgcaga actggtaggt atggaagatc cctcgagatc
301 cattgtgctg gcggtaggcg agcagcgcct gcctgaagct gcgggcattc ccgatcagaa
361 atgagcgcca gtcgtcgtcg gctctcggca ccgaatgcgt atgattctcc gccagcatgg
421 cttcggccag tgcgtcgagc agcgcccgct tgttcctgaa gtgccagtaa agcgccggct
481 gctgaacccc caaccgttcc gccagtttgc gtgtcgtcag accgtctacg ccgacctcgt
541 tcaacaggtc cagggcggca cggatcactg tattcggctg caactttgtc atgcttgaca
601 ctttatcact gataaacata atatgtccac caacttatca gtgataaaga atccgcgcca
661 gcacaatgga tctcgaggtc gagggatctc tagaggatcc tctacgccgg acgcatcgtg
721 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
781 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
841 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
901 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
961 aagggagagc gtcgacctcg ggccgcgttg ctggcgtttt tccataggct ccgcccccct
1021 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
1081 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
1141 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
1201 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
1261 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
1321 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
1381 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
1441 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
1501 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
1561 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
1621 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
1681 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
1741 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
1801 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
1861 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
1921 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
1981 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
2041 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
2101 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
2161 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
2221 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
2281 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
2341 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
2401 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
2461 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
2521 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
2581 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
2641 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
2701 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
2761 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctgatggctc
2821 tttgcggcac ccatcgttcg taatgttccg tggcaccgag gacaaccctc aagagaaaat
2881 gtaatcacac tggctcacct tcgggtgggc ctttctgcgt ttataaggag acactttatg
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