請在線咨詢或直接聯系18986294316/17720555249

必騰預構載體定制最高100元/個

基本信息

載體描述 

注意：

客戶在我公司選擇構建載體服務項目時，請自行提供母源載體。

如要使用本公司保存的母源載體時，本公司不能保證所有母源載體完全符合客戶預期，也不承擔由此產生的任何責任。

由質粒pCold-I改造，在Ndel與Sacl酶切位點間，插入ZZdomain：

Z Zdomain：

GACAACAAATTCAACAAAGAACAACAAAACGCGTTCTATGAGATCTTACATTTACCTAACTTAAACGAAGAACAACGAAACGCCTTCATCCAAAGTTTAAAAGATGACCCAAGCCAAAGCGCTAACCTTTTAGCAGAAGCTAAAAAGCTAAATGATGCTCAGGCGCCGAAAGTAGACAACAAATTCAACAAAGAACAACAAAACGCGTTCTATGAGATCTTACATTTACCTAACTTAAACGAAGAACAACGAAACGCCTTCATCCAAAGTTTAAAAGATGACCCAAGCCAAAGCGCTAACCTTTTAGCAGAAGCTAAAAAGCTAAATGATGCTCAGGCGCCGAAAGTA

ZZ domain總長348bp，由兩段完全一樣的序列組成，改造后為TEE+His+Factor Xa+ZZ+多克隆位點。

載體序列

LOCUS       Exported                4788 bp ds-DNA     circular SYN 16-MAR-2019

DEFINITION  .

ACCESSION   .

VERSION     .

KEYWORDS    pCold-ZZ

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4788)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Saturday, March 16, 2019 from SnapGene 3.2.1

FEATURES             Location/Qualifiers

     source          1..4788

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        15..81

                     /gene="cspA"

                     /note="cspA promoter"

                     /note="promoter of the E. coli cold shock protein cspA gene

                     (Mitta et al., 1997)"

                     /note="color: #ffffff; direction: RIGHT"

     protein_bind    84..100

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="lac operator"

                     /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)."

                     /note="color: #31849b"

     5'UTR           122..255

                     /gene="cspA"

                     /note="cspA 5'UTR"

                     /note="5'UTR of the E. coli cold shock protein cspA gene

                     (Mitta et al., 1997)"

                     /note="color: #00ff00"

     CDS             256..270

                     /codon_start=1

                     /gene="cspA"

                     /product="translation enhancing element for E. coli (Qing

                     et al., 2004)"

                     /note="TEE"

                     /note="color: #ff00ff"

                     /translation="MNHKV"

     CDS             271..288

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /note="6xHis"

                     /note="color: #ff9900"

                     /translation="HHHHHH"

     CDS             289..300

                     /codon_start=1

                     /product="Factor Xa recognition and cleavage site"

                     /note="Factor Xa site"

                     /note="color: #3366ff

                     Cleavage site after base 300"

                     /translation="IEGR"

     CDS             328..498

                     /codon_start=1

                     /product="IgG-binding unit of Staphylococcus aureus protein

                     A"

                     /note="ProtA"

                     /note="color: #ccffcc"

                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA

                     EAKKLNDAQAPK"

     CDS             502..672

                     /codon_start=1

                     /product="IgG-binding unit of Staphylococcus aureus protein

                     A"

                     /note="ProtA"

                     /note="color: #ccffcc"

                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA

                     EAKKLNDAQAPK"

     misc_feature    688..741

                     /note="MCS"

                     /note="Multi clone site"

                     /note="color: #00ccff"

     3'UTR           749..893

                     /gene="cspA"

                     /note="cspA 3'UTR"

                     /note="3'UTR of the E. coli cold shock protein cspA gene

                     (Mitta et al., 1997)"

                     /note="color: #00ff00"

     rep_origin      complement(1092..1547)

                     /direction=LEFT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

                     /note="color: #ffff00"

     promoter        1647..1751

                     /gene="bla"

                     /note="AmpR promoter"

                     /note="color: #ffffff; direction: RIGHT"

     CDS             1752..2612

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /note="AmpR"

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /note="This feature has 2 segments:

                        1: 1752 .. 1820 / #ccffcc / signal sequence

                        2: 1821 .. 2612 / #ccffcc

                     Cleavage site after base 1820"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      2783..3371

                     /direction=RIGHT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

                     replication"

                     /note="color: #ffff00"

     CDS             complement(3543..4625)

                     /codon_start=1

                     /gene="lacI"

                     /product="lac repressor"

                     /note="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)."

                     /note="color: #993366"

                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL

                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV

                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH

                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA

                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC

                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR

                     ALADSLMQLARQVSRLESGQ"

     promoter        complement(4626..4703)

                     /gene="lacI"

                     /note="lacI promoter"

                     /note="color: #ffffff; direction: LEFT"

ORIGIN

        1 aaggaatggt gtggccgatt aatcataaat atgaaaaata attgttgcat cacccgccaa

       61 tgcgtggctt aatgcacatc aaattgtgag cggataacaa tttgatgtgc tagcgcatat

      121 ccagtgtagt aaggcaagtc ccttcaagag ttatcgttga tacccctcgt agtgcacatt

      181 cctttaacgc ttcaaaatct gtaaagcacg ccatatcgcc gaaaggcaca cttaattatt

      241 aagaggtaat acaccatgaa tcacaaagtg catcatcatc atcatcatat cgaaggtagg

      301 catatggcgc aacacgatga agccgtagac aacaaattca acaaagaaca acaaaacgcg

      361 ttctatgaga tcttacattt acctaactta aacgaagaac aacgaaacgc cttcatccaa

      421 agtttaaaag atgacccaag ccaaagcgct aaccttttag cagaagctaa aaagctaaat

      481 gatgctcagg cgccgaaagt agacaacaaa ttcaacaaag aacaacaaaa cgcgttctat

      541 gagatcttac atttacctaa cttaaacgaa gaacaacgaa acgccttcat ccaaagttta

      601 aaagatgacc caagccaaag cgctaacctt ttagcagaag ctaaaaagct aaatgatgct

      661 caggcgccga aagtagacgc gacattcgag ctcggtaccc tcgagggatc cgaattcaag

      721 cttgtcgacc tgcagtctag ataggtaatc tctgcttaaa agcacagaat ctaagatccc

      781 tgccatttgg cggggatttt tttatttgtt ttcaggaaat aaataatcga tcgcgtaata

      841 aaatctatta ttatttttgt gaagaataaa tttgggtgca atgagaatgc gcaggccctt

      901 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac

      961 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc

     1021 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag

     1081 agtgcaccat aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta

     1141 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga

     1201 atagcccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa

     1261 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga

     1321 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc

     1381 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga

     1441 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg

     1501 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt

     1561 tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgt

     1621 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac

     1681 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa

     1741 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat

     1801 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc

     1861 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga

     1921 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg

     1981 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc

     2041 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag

     2101 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc

     2161 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg

     2221 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg

     2281 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac

     2341 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac

     2401 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg

     2461 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg

     2521 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg

     2581 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac

     2641 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg

     2701 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg

     2761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc

     2821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc

     2881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt

     2941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc

     3001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact

     3061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac

     3121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag

     3181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg

     3241 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg

     3301 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga

     3361 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt

     3421 ttgctcacat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca

     3481 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc

     3541 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc

     3601 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt

     3661 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg

     3721 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata

     3781 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc

     3841 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc

     3901 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg

     3961 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta

     4021 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg

     4081 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg

     4141 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca

     4201 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc

     4261 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg

     4321 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc

     4381 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc

     4441 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc

     4501 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg

     4561 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt

     4621 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag

     4681 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat

     4741 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgc

//