Plasmid classification: lactation cell plasmid; lactation expression plasmid; pCAG series plasmid.
Cloning strains: E. coli DH5 and E.
LOCUS Exported 6200 bp ds-DNA circular SYN 18-AUG-2017
EYWORDS pCIG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6200)
FEATURES Location/Qualifiers
source 1..6200
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(42..630)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(801..1661)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1662..1766)
/gene="bla"
/note="AmpR promoter"
enhancer 1797..2176
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2178..2455
/note="chicken beta-actin promoter"
intron 2456..3468
/note="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
misc_feature 3594..4172
/note="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4173..4892
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_site 5049..5104
/note="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal"
promoter 5668..5863
/note="SV40 promoter"
/note="SV40 early promoter"
rep_origin 5714..5849
/note="SV40 ori"
/note="SV40 origin of replication"
polyA_signal 5869..6003
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
ORIGIN
1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
481 agnacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
2161 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
2221 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
2281 ggcggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg
2341 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
2401 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
2461 cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct
2521 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
2581 taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa
2641 agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg
2701 tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg
2761 cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc
2821 ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
2881 ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc
2941 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
3001 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
3061 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
3121 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
3181 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
3241 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
3301 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
3361 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
3421 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
3481 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tgctcgaggt cgatcgacgg
3541 tatcgataag cttgatatcg aattcctgca gcccggccgc tacgtaaatt ccgccccccc
3601 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
3661 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
3721 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
3781 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
3841 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
3901 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
3961 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
4021 ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg
4081 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
4141 ttgaaaaaca cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca
4201 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
4261 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
4321 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
4381 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
4441 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
4501 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
4561 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
4621 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
4681 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
4741 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
4801 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
4861 tcactctcgg catggacgag ctgtacaagt aaagcggccg caattcactc ctcaggtgca
4921 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
4981 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
5041 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
5101 atcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
5161 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
5221 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
5281 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
5341 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
5401 catagctgtc cctcttctct tatggagatc cctcgacctg caccgtcgac cagctggtcg
5461 acggtgcacc gtcgaccagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
5521 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
5581 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
5641 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
5701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
5761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
5821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
5881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
5941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
6001 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
6061 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
6121 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
6181 acatgtgagc aaaaggccag
//
Caution:
1. This product is FOR RESEARCH USE ONLY!
2. The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.
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