pComb3XSS

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载体基本信息

质粒名称: pComb3XSS 
出品公司: 爱迪基因
目录编号: 63890
存储实验室: Carlos Barbas

载体骨架基本信息
载体名称: pComb3H
载体抗性: 氨苄霉素
空载体大小: 3300 bp
完整质粒大小: 4900 bp
载体修饰: Addition of 6xHis and HA tags for detection and purification, amber stop codon upstream of geneIII to produce soluble protein by switching to a non-suppressor strain of E. coli (e.g. Top10F')
载体类型: 细菌表达;phage display
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: SS Stuffer
物种来源: Synthetic
插入大小: 1600 bp
基因又名: SfiI Stuffer
启动子 lacZ
标签/融合蛋白:
6x His (C terminal on backbone)
HA tag (C terminal on backbone)
geneIII (C terminal on backbone)
克隆方法: 限制性内切酶
5' 克隆位点: SfiI(未破坏)
3' 克隆位点: SfiI (未破坏)
5' 测序引物 5'-AAG ACA GCT ATC GCG ATT GCA G-3'
3' 测序引物 5'-GCC CCC TTA TTA GCG TTT GCC ATC-3'

文献引用方法
材料和方法部分: pComb3XSS was a gift from Carlos Barbas (爱迪基因 plasmid # 63890)
参考文献部分: Methods for the generation of chicken monoclonal antibody fragments by phage display. Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF 3rd. J Immunol Methods. 2000 Aug 28;242(1-2):159-81. S0022175900002210 [pii] PubMed 10986398

载体图谱质粒图谱和多克隆位点信息

pComb3XSS质粒图谱

载体简介

载体序列

LOCUS       pComb3XSS                4991 bp ds-DNA     circular SYN 10-JAN-2018
DEFINITION  3rd generation plasmid for phage display on modified geneIII, 
            contains stuffer fragment.
ACCESSION   .
VERSION     .
KEYWORDS    pComb3XSS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  AUTHORS   Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF 3rd
  TITLE     Methods for the generation of chicken monoclonal antibody fragments 
            by phage display.
  JOURNAL   J Immunol Methods. 2000 Aug 28;242(1-2):159-81.
  PUBMED    10986398
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|pComb3XSS|
FEATURES             Location/Qualifiers
     source          1..4991
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    109..130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        145..175
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    183..199
                     /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)."
     primer_bind     complement(406..422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(406..422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(419..441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    430..446
                     /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(454..484)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(574..591)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1584..1907
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANL"
     CDS             1962..1979
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1986..2012
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      complement(2606..3061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2743..2764)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2955..2974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3088..3192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3193..4053
                     /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     complement(3411..3430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4224..4812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4713..4732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4966..4983
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
       61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      181 aattgtgagc ggataacaat tgaattcagg aggaatttaa aatgaaaaag acagctatcg
      241 cgattgcagt ggcactggct ggtttcgcta ccgtggccca ggcggccgag ctcgccatgg
      301 ctggttgggc agcgagtaat aacaatccag cggctgccgt aggcaatagg tatttcatta
      361 tgactgtctc cttggcgact agctagttta gaattcgtaa tcatggtcat agctgtttcc
      421 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      481 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      541 cgctttccag tcgggaaacc tgtcgtgtta ctaatgatgg tgatggtgat ggctagtttt
      601 gtcacaagat ttgggctcaa ctttcttgtc caccttggtg ttgctgggct tgtgattcac
      661 gttgcagatg taggtctggg tgcccaagct gctggagggc acggtcacca cgctgctgag
      721 ggagtagagt cctgaggact gtaggacagc cgggaaggtg tgcacgccgc tggtcagggc
      781 gcctgagttc cacgacaccg tcgccggttc ggggaagtag tccttgacca ggcagcccag
      841 ggccgctgtg cccccagagg tgctcttgga ggagggtgcc agggggaaga ccgatgggcc
      901 cttggtggag gctgcggaga cggtgaccgt ggtaccagca gaaacctggc caggctccca
      961 ggctcctcat ctatggtaca tccagcaggg ccactggcat cccagacagg ttcagtggca
     1021 gtgggtctgg gacagacttc actctcacca tcagcagact ggagcctgaa gattttgcag
     1081 tgtactactg tcagcagtat ggtggctcac cgtggttcgg ccaagggacc aaggtggaac
     1141 tcaaacgaac tgtggctgca ccatctgtct tcatcttccc gccatctgat gagcagttga
     1201 aatctggaac tgcctctgtt gtgtgcctgc tgaataactt ctatcccaga gaggccaaag
     1261 tacagtggaa ggtggataac gccctccaat cgggtaactc ccaggagagt gtcacagagc
     1321 aggacagcaa ggacagcacc tacagcctca gcagcaccct gacgctgagc aaagcagact
     1381 acgagaaaca caaagtctac gcctgcgaag tcacccatca gggcctgagc ttgcccgtca
     1441 caaagagctt caacagggga gagtgttagt tctagataat taattaggag gaatttaaaa
     1501 tgaaatacct attgcctacg gcagccgctg gattgttatt actcgctgcc caaccagcca
     1561 tggccgaggt gcagctgctc gagatgagcg ataaaattat tcacctgact gacgacagtt
     1621 ttgacacgga tgtactcaaa gcggacgggg cgatcctcgt cgatttctgg gcagagtggt
     1681 gcggtccgtg caaaatgatc gccccgattc tggatgaaat cgctgacgaa tatcagggca
     1741 aactgaccgt tgcaaaactg aacatcgatc aaaaccctgg cactgcgccg aaatatggca
     1801 tccgtggtat cccgactctg ctgctgttca aaaacggtga agtggcggca accaaagtgg
     1861 gtgcactgtc taaaggtcag ttgaaagagt tcctcgacgc taacctggcg tacccgtacg
     1921 acgttccgga ctacggttct actagtggcc aggccggcca gcaccatcac catcaccatg
     1981 gcgcataccc gtacgacgtt ccggactacg cttcttagga gggtggtggc tctgagggtg
     2041 gcggttctga gggtggcggc tctgagggag gcggttccgg tggtggctct ggttccggtg
     2101 attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa aatgccgatg
     2161 aaaacgtgct acagtctgac gctaaaggca aacttgattc tgtcgctact gattacggtg
     2221 ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat ggtgctactg
     2281 gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt
     2341 taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa tgtcgccctt
     2401 ttgtctttgg cgctggtaaa ccatatgaat tttctattga ttgtgacaaa ataaacttat
     2461 tccgtggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta ttttctacgt
     2521 ttgctaacat actgcgtaat aaggagtctt aagctagcta attaatttaa gcggccgcag
     2581 atctgctctc tgaggaggat ctgggaaatt gtaagcgtta atattttgtt aaaattcgcg
     2641 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2701 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2761 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2821 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2881 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2941 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3001 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3061 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3121 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3181 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3241 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3301 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3361 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3421 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3481 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3541 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3601 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3661 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3721 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3781 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3841 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3901 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3961 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4021 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4081 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4141 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4201 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4261 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4321 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4381 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4441 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4501 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4561 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4621 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4681 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4741 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4801 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4861 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4921 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4981 gaggaagcgg a
//

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