pEGM-7ZF(+)

pEGM-7ZF(+)

载体基本信息

出品公司:	Ybscience
载体名称:	pEGM-7ZF(+)
质粒类型:	大肠杆菌克隆载体
高拷贝/低拷贝:	--
启动子:	--
克隆方法:	多克隆位点，限制性内切酶
载体大小:	3000bp
5' 测序引物及序列:	--
3' 测序引物及序列:	--
载体标签:	--
载体抗性:	Ampicillin
筛选标记:	--
备注:	--
稳定性:	--
组成型:	--
病毒/非病毒:	非病毒

 

载体质粒图谱和多克隆位点信息

载体简介

 

The pGEM?-7Zf(+) Vector(a) is a derivative of the pGEM?-3Zf(+) Vector(a) and contains
the origin of replication of the filamentous phage f1. The plasmid serves as a
standard cloning vector, as a template for in vitro transcription, and it can be used
for the production of circular ssDNA. The plasmid contains SP6 and T7 RNA polymerase
promoters flanking a region of multiple cloning sites within the α-peptide
coding region of β-galactosidase (1). Insertional inactivation of the α-peptide allows
recombinant clones to be directly identified by color screening on indicator plates.
The multiple cloning region is unique and includes restriction sites for Apa I, Aat II,
Sph I, Xba I, Xho I, EcoR I, Kpn I, Sma I, Csp45 I, Cla I, Hind III, BamH I, Sac I,
BstX I and Nsi I. This arrangement is designed specifically for generation of unidirectional
deletions with the Erase-a-Base? System(b). The polylinker contains
restriction enzyme sites that produce 5′ overhangs or blunt ends (sensitive to
Exonuclease III) flanked on both sides by blocks of restriction sites that generate
3′ overhangs (resistant to Exonuclease III).
For induction of ssDNA, bacterial cells containing pGEM?-7Zf(+) recombinants are
infected with an appropriate helper phage. The plasmid then enters the f1 replication
mode, and the resulting ssDNA is exported from the cell as an encapsidated viruslike
particle. The sequence of the ssDNA rescued upon infection with helper phage
is complementary to the sequence shown in Figure 1. The exported ssDNA can be
used for mutagenesis in vitro or can be sequenced using the T7 Promoter Primer
(Cat.# Q5021) or pUC/M13 Forward Primer (Cat.# Q5391, Q5601).

 

载体序列

LOCUS       pGEM-7Zf-	3000 bp 	DNA	SYN
DEFINITION  pGEM-7Zf-
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..3000
                     /organism="pGEM-7Zf-"
                     /mol_type="other DNA"
     misc_feature    complement(123..140)
                     /label="Sp6_primer"
     promoter        complement(154..172)
                     /label="M13_reverse_primer"
     misc_feature    complement(171..193)
                     /label="M13_pUC_rev_primer"
     promoter        complement(207..236)
                     /label="lac_promoter"
     rep_origin      complement(545..1164)
                     /label="pBR322_origin"
     gene            complement(1319..2179)
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             complement(1319..2179)
                     /label="ORF frame 3"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        complement(2221..2249)
                     /label="AmpR_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     rep_origin      2382..2688
                     /label="f1_origin"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    complement(2832..2980)
                     /label="lacZ_a"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    2946..2968
                     /label="M13_pUC_fwd_primer"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        2961..2977
                     /label="M13_forward20_primer"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        2984..2
                     /label="T7_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
ORIGIN
    1 gggcgaattg ggcccgacgt cgcatgctcc tctagactcg aggaattcgg taccccgggt
   61 tcgaaatcga taagcttgga tccggagagc tcccaacgcg ttggatgcat agcttgagta
  121 ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
  181 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
  241 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
  301 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
  361 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
  421 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
  481 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
  541 aggccgcgtt gctggcgttt ttcgataggc tccgcccccc tgacgagcat cacaaaaatc
  601 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
  661 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
  721 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
  781 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
  841 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
  901 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
  961 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
 1021 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
 1081 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
 1141 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
 1201 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
 1261 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
 1321 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
 1381 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
 1441 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
 1501 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
 1561 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
 1621 ttgttggcat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
 1681 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
 1741 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
 1801 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
 1861 tgactggtga gtactcaacc aagtcattct gagaataccg cgcccggcga ccgagttgct
 1921 cttgcccggc gtcaatacgg gataatagtg tatgacatag cagaacttta aaagtgctca
 1981 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
 2041 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
 2101 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
 2161 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
 2221 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
 2281 cgcgcacatt tccccgaaaa gtgccacctg tatgcggtgt gaaataccgc acagatgcgt
 2341 aaggagaaaa taccgcatca ggcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
 2401 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
 2461 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
 2521 ggctcccttt agggttccga tttagagctt tacggcacct cgaccgcaaa aaacttgatt
 2581 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
 2641 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
 2701 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
 2761 atgagctgat ttaacaaata tttaacgcga attttaacaa aatattaacg tttacaattt
 2821 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
 2881 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
 2941 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata