| 质粒名称: | PB-CA sfGFP |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 159094 |
| 存储实验室: | Mario Capecchi |
|
载体骨架基本信息 |
|
| 载体名称: | PB-CA |
| 载体出品公司: | Andras Nagy |
| 完整质粒大小: | 7274 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | DH10B |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sfGFP |
| 物种来源: | Synthetic |
|
文献引用方法 |
|
| 材料和方法部分: | PB-CA sfGFP was a gift from Mario Capecchi (Addgene plasmid # 159094 ; http://n2t.net/addgene:159094 ; RRID:Addgene_159094) |
买家导航
PB-CA sfGFP载体质粒基本信息
PB-CA sfGFP质粒图谱载体图谱和PB-CA sfGFP载体序列质粒序列多克隆位点信息
PB-CA sfGFP质粒载体简介
PB-CA sfGFP质粒序列载体序列
LOCUS PB-CA sfGFP 7274 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION PB-CA sfGFP piggyBac transposon.
ACCESSION .
VERSION .
KEYWORDS PB-CA sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7274)
AUTHORS Nam HS, Capecchi MR
TITLE piggyBac transposon plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7274)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7274
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..659
/label=chicken beta-actin promoter
intron 660..1676
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1600..1622
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1684..1703
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1745..2458
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind complement(1775..1803)
/label=GFP-R
/note="GFP, reverse primer. Does NOT anneal to EGFP"
primer_bind 2398..2418
/label=GFP-F
/note="GFP, forward primer. Does NOT anneal to EGFP"
misc_feature 2498..3086
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2551..2571)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2969..2980)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(3125..3142)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3131..3355
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 3780..3814
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
promoter complement(3963..3981)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4002..4018)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4002..4018)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4015..4037)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4026..4042
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4050..4080)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4095..4116
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4233..4250)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4404..4992)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4484..4503)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5163..6023)
/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"
primer_bind 5786..5805
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6024..6128)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6154..6609)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6291..6312)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6503..6522
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6736..6758
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6750..6767
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6751..6767
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6774..6793
/label=T7
/note="T7 promoter, forward primer"
promoter 6774..6792
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
repeat_region 6859..6921
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
481 tgggggcggg gggggggggg gggcgcgcgc cggggggggg gggggggggs ggggggsggg
541 gsggggsgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
1261 ggccgcctcg ggccggggrg ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg ctagcgccgc
1741 caccatgagc aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
1801 tggtgatgtt aatgggcaca aattttctgt ccgtggagag ggtgaaggtg atgctacaaa
1861 cggaaaactc acccttaaat ttatttgcac tactggaaaa ctacctgttc cgtggccaac
1921 acttgtcact actctgacct atggtgttca atgcttttcc cgttatccgg atcacatgaa
1981 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc
2041 tttcaaagat gacgggacct acaagacgcg tgctgaagtc aagtttgaag gtgataccct
2101 tgttaatcgt atcgagttaa agggtattga ttttaaagaa gatggaaaca ttcttggaca
2161 caaactcgag tacaacttta actcacacaa tgtatacatc acggcagaca aacaaaagaa
2221 tggaatcaaa gctaacttca aaattcgcca caacgttgaa gatggttccg ttcaactagc
2281 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
2341 ttacctgtcg acacaatctg tcctttcgaa agatcccaac gaaaagcgtg accacatggt
2401 ccttcttgag tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaatg
2461 attaattaat gatcatacag tgctctgagt acgctataat caacctctgg attacaaaat
2521 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
2581 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
2641 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
2701 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
2761 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
2821 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
2881 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
2941 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
3001 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
3061 gatctccctt tgggccgcct ccccgcagga tccactagag cggccgccta gagctcgcta
3121 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
3181 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
3241 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
3301 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
3361 gaggcggaaa gaaccagctg gggctcgact agagcttgcg gaacccttaa gcttctagcc
3421 aacaagctcg tcatcgcttt gcagaagagc agagaggata tgctcatcgt ctaaagaact
3481 acccatttta ttatatatta gtcacgatat ctataacaag aaaatatata tataataagt
3541 tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag
3601 tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag
3661 tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc
3721 ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc
3781 atgcgtcaat tttacgcaga ctatctttct agggttaaaa aagatttgcg ctttactcga
3841 cctaaacttt aaacacgtca tagaatcttc gtttgacaaa aaccacattg tggccaagct
3901 gtgtgacgcg acgcgcgcta aagaatggca aaccaagtcg cgcgaggtac cagcttttgt
3961 tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg
4021 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
4081 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
4141 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
4201 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
4261 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
4321 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
4381 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
4441 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
4501 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
4561 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
4621 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
4681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
4741 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
4801 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
4861 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
4921 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
4981 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
5041 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
5101 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
5161 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
5221 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
5281 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
5341 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
5401 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
5461 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
5521 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
5581 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
5641 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
5701 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
5761 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
5821 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
5881 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
5941 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
6001 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
6061 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
6121 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
6181 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
6241 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
6301 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
6361 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
6421 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
6481 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
6541 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
6601 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
6661 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
6721 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
6781 actcactata gggcgaattg gagctcggta ttcacgacag caggctgaat aataaaaaaa
6841 ttagaaacta ttatttaacc ctagaaagat aatcatattg tgacgtacgt taaagataat
6901 catgcgtaaa attgacgcat gtgttttatc ggtctgtata tcgaggttta tttattaatt
6961 tgaatagata ttaagtttta ttatatttac acttacatac taataataaa ttcaacaaac
7021 aatttattta tgtttattta tttattaaaa aaaaacaaaa actcaaaatt tcttctataa
7081 agtaacaaaa cttttaaaca ttctctcttt tacaaaaata aacttatttt gtactttaaa
7141 aacagtcatg ttgtattata aaataagtaa ttagcttaac ctatacataa tagaaacaaa
7201 ttatacttat tagtcagtca gaaacaactt tggcacatat caatattatg ctctcgttaa
7261 taccggtcat ggtc
//
