| 质粒名称: | pOBCol2.3-GFPemd |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 110210 |
| 存储实验室: | David Rowe |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2700 bp |
| 完整质粒大小: | 7500 bp |
| 载体修饰: | MCS中的Hind3和EcoR1位点通过适配器转换成Sal1位点。 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | eGFPemerald,Emerald |
| 物种来源: | A. victoria |
| 插入大小: | 720 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | 2.3 kb rat type 1 collagen + first intron |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Hind3（克隆过程中销毁） |
| 3' 克隆位点: | EcoR1（克隆过程中销毁） |
| 5' 测序引物 | M13reverse |
| 3' 测序引物 | M13forward |
|
文献引用方法 |
|
| 材料和方法部分: | pOBCol2.3-GFPemd was a gift from David Rowe (Addgene plasmid # 110210 ; http://n2t.net/addgene:110210 ; RRID:Addgene_110210) |
| 参考文献部分: | Use of type I collagen green fluorescent protein transgenes to identify subpopulations of cells at different stages of the osteoblast lineage. Kalajzic I, Kalajzic Z, Kaliterna M, Gronowicz G, Clark SH, Lichtler AC, Rowe D. J Bone Miner Res. 2002 Jan;17(1):15-25. doi: 10.1359/jbmr.2002.17.1.15. 10.1359/jbmr.2002.17.1.15 PubMed 11771662 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pOBCol2.3-GFPemd 7627 bp ds-DNA circular SYN 07-AUG-2018
DEFINITION Labels cells in the late osteoblast lineage..
ACCESSION .
VERSION .
KEYWORDS pOBCol2.3-GFPemd
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7627)
AUTHORS Kalajzic I, Kalajzic Z, Kaliterna M, Gronowicz G, Clark SH, Lichtler
AC, Rowe D
TITLE Use of type I collagen green fluorescent protein transgenes to
identify subpopulations of cells at different stages of the
osteoblast lineage.
JOURNAL J Bone Miner Res. 2002 Jan;17(1):15-25. doi:
10.1359/jbmr.2002.17.1.15.
PUBMED 11771662
REFERENCE 2 (bases 1 to 7627)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 7, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7627
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 10..32
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 24..41
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 25..41
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 4125..4134
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4131..4850
/codon_start=1
/product="Emerald GFP"
/label=EmGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4176..4197)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4437..4456)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4784..4805
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 4883..5017
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4920..4939)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4974..4993
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5052..5068)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5052..5068)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5065..5087)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5076..5092
/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(5100..5130)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5145..5166
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5283..5300)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5454..6042)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5534..5553)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6213..7073)
/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 6836..6855
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7074..7178)
/gene="bla"
/label=AmpR promoter
primer_bind 7246..7264
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7302..7324)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7424..7443
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg cgtcgacgca
61 gcttggaacc cactgcccac aactcctggg gcaaaaacct ggagtcagac atggggtgaa
121 ggctgtcaca cggcacagac acgtcaagca ccccccccaa ttctagtagt ctcctagcct
181 ccaccagaac cccagaccct tgatgtggca gtcaccagtc cacacctgtt aggctcttgt
241 ctcttcttcc agatgagcct ggggggcggg ggggtgctag atcaggagca gggaaaagta
301 gctttggata agtgcttttc ccaatacaaa acccaacaaa gagtgggcag atcacactgt
361 gtagtgcttc gtggaaccct accctagaca actgccttga acacctattc cctctgatgt
421 acaccatccc cgtccactgt tagggagtgg gcatcctttg gaactgacca ctgtggaagg
481 caggacttta ctgagttccg gaactaccat ctcagcttct cagccccagc cttaccctac
541 aggcactggc ataggcgggg gcagatcctg ggccacaagt cactgccaca tggttgggat
601 aattgatgaa gtcctgtcct tccattgctg tctccagttc tgcttctctg gaaactctat
661 atttttccct ttaattatag cctctgcagt ctccctctgc caccccaccc gcaccgctta
721 gcctaactgc ccacggccag cgacgtggct ccctcccctt ctgctccctt ggtctttttt
781 attttttttt ctttgccttc gttgcacaaa actagctcag ggagggcgtg aagggggggg
841 gagcaatgga atcttggatg gtttggagga ggcgggactc cttgcttcca cgtttacagc
901 tctgaagacg gctgtggggg aagtgataca ggacgtctat gggccctgag aggagacccc
961 tatgcttccc tgccacccac acagtttaac aaaatgaagt tcctaagtag agtgggggtc
1021 aggcagagca cctttgcagg gttgatggga gcccagggaa agaaaggaca ctgtctttta
1081 gggacacatt taaatataag ccacttttct tgggggacga caaatgaccc tttcctgatt
1141 gcagaggtgg ggaacaatgg ctgagatttt cagcaaagaa gcgaggacat gaggagtagc
1201 cttcaaataa agtcactcag ctaccaaaaa caagtttctg ccacacaccg agttcaccta
1261 ggtgtcccca gaccagatcc aagtacagta aggaaagcag gttctctaca gagagaacac
1321 ggctctatgg ccaatgcctt ctacctgctc tttctggatt gatactgcta cctaagaggg
1381 cctctaacca attcctggct gtagccacag ctgacacaag acctttttct aagacatccc
1441 tggtcacagg cctcctgtag caaattccag ccctgggatg gaggtggtca ggaaagagtt
1501 tatacaagaa gacccaggcc acagctttaa ggactcagaa acccccctgc ccacacggct
1561 gcccatcata acgcagaagg tttcttctgg aaggacaagg atgtcaaact tctccccaag
1621 cctaatcctc agagatgtct ccctctgtta cacctggggc tggagaaagg tgggtctttc
1681 atggagccac attcatggca gaacagatag ccaccccact cctttcaaac aaccacatat
1741 ctgactctta gtatctgtga agagatgtct aatttgttcc caaatattcc taccctgcat
1801 acctgggccc acaccatgag gtattctcct ccctctaaca gtcacatctg cttagctgcc
1861 tggttcttcg gatttggaga gatgcttgcc taacttattc ttccttaggt cttcccaagg
1921 atgccagaaa gactatgaga catggccaag aggacctttt cccaattgtg cctgacactg
1981 aaccctttgt aatgttcccc aactcagatt cccaattcta catccttctg atttgaggtc
2041 ccagaaggaa agtgcaaggg gcatccccta cccacaatca gtatatcgag ggcccagcca
2101 cactcagtga tagcacctct ggcccatgta gatctggggg acaagggtgg cagaattgca
2161 aaggggggag ggggctgggt ggactccttt cccttccttt ccctcctccc ccctcttcgt
2221 tccaaattgg gggccgggcc aggcagttct gattggctgg gggccgggct gctggctccc
2281 cctctccaag aggcagggtt cctcccagcc ctcctccatc aggatggtat aaaaggggcc
2341 caggccagtc gtcggagcag acgggagttt cacctccgga cggagcagga ggcacacgga
2401 gtgaggccac gcatgagccg aagctaaccc cccaccccag ccgcaaagag tctacatgtc
2461 tagggtctag aggcggccgc cttcagcttt gtggacctcc ggctcctgct cctcttaggg
2521 gccactgccc tcctgacgca tggccaagaa gacagtaagt tccaaactct ggggaggtta
2581 tggtttcttg ctatcgcgtt ctgcataggg gtcctagcaa taaagactta aacggaagcg
2641 cggggtgctc cagagttagg agggatgaga aggtgatgga gaggaactgg gaagagagag
2701 gtcaccctcg cgagcagagg agagtgagcg gatctcagaa gtggagccct ggttgagatg
2761 gcacggcagg aaatttcaag catctatctg tagcgcgacg ggaaaagtgg gaatctgggc
2821 aggggttggg tcagagtacg tcgaagatca aataagacct ctggaataga gattcgaagc
2881 tcaagatgta aaagcaatca aaaaggctgc gggatgattc ataaggaaag attgtttgcc
2941 ctctctgcag gctagacagt gttgctgggg cctccggggt gctgggctct gggagtgggg
3001 tggggggagg aagggacgca cggagagggc gggttgctgg atgagaaagt ttggctaaga
3061 cttgactggg cgcggtccct gtgcctcctg ctggtcaacc ttgagcacta cgagtcctgg
3121 tacagtactt tacagcgcgc gggcttaaga gggaaaacct gggctattgc cgggcctgga
3181 ccctcatctc cagactagga aaataaaagc acagatggat gtgatagctt tggtctttga
3241 aagtgaccca ggccagtaaa attgggtccg ccccgtgcat tgcttagcgt tgcccgccac
3301 ctagtggttg ctttgatccg gtgctagaga ggtgggcgtg gttaactaat cattgctatt
3361 tgtagacttg agttctttct ttggctaagg agtgacccca aagctctact ggctttgaag
3421 gaaggaccgg gcaagctttg attcctggga gcggagagtt agggtgtatc taactttagg
3481 gatatatttt actactctgg acaaggctcc acggaagtca ccctcctcca ttttagcccc
3541 gcccattccg tcccacctgc ccaaggaaag gaggtgggac cagggctgga atttcccttc
3601 tctcctccca cccccgcctc cggtctcccc gcaaaccacg tcccacaaac ttccacctac
3661 tttgtaggag ccctcctact ctccgtcagg aggacttcta tttcatttgg ttcccgcgcg
3721 cgtgggctga agttgagttt ggaagcaacc ccagtctaaa gctttgttta aattcctggg
3781 aacttggaag taacagccct ggccaggctt cttggtgctg tcactgaacg ccgttacaga
3841 gcagacctgc atttaaggat ttaagggaag aaaatagggg ttgggggaag atgtgggaag
3901 tggaaaatct acgtgcacct cttagacagg ggaaagggtg agtaaggctg ggatccggtg
3961 gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg
4021 gaagtgttac ttctgctcta aaagctgctg caggtcttag ttaactaaga ctctagggaa
4081 gatcccccat caagctagct tggcattccg gtactgttgg taaagccacc atggtgagca
4141 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
4201 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
4261 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
4321 ccttgaccta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
4381 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
4441 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
4501 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
4561 acaactacaa cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
4621 tgaacttcaa gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
4681 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
4741 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
4801 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa ctcgactaga
4861 gtcggggcgg ccgacctcga ggaacttgtt tattgcagct tataatggtt acaaataaag
4921 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
4981 gtccaaactc atcaatgtat cttatcatgt ctggatcgat cccccgaatt ggtcgaccaa
5041 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
5101 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
5161 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
5221 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
5281 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
5341 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
5401 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
5461 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
5521 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
5581 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
5641 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
5701 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
5761 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
5821 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
5881 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
5941 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
6001 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
6061 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
6121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
6181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
6241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
6301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
6361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
6421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
6481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
6541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
6601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
6661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
6721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
6781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
6841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
6901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
6961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
7021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
7081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
7141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
7201 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
7261 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
7321 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
7381 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
7441 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
7501 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
7561 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
7621 gtaacgc
//
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