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载体序列
LOCUS pQlinkN 4823 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pQLinkN
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4823)
AUTHORS Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K
TITLE Vectors for co-expression of an unrestricted number of proteins.
JOURNAL Nucleic Acids Res. 2007 Feb 20. ():.
PUBMED 17311810
REFERENCE 2 (bases 1 to 4823)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4823
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 36..80
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 56..72
/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)."
protein_bind 88..104
/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)."
RBS 123..134
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
protein_bind complement(184..208)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
terminator 230..324
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 418..1077
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(484..503)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 1142..1228
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1321..2403)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 2365..2384
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(2404..2481)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
primer_bind complement(2565..2587)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 2673..2813
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 2728..2747
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(2828..2845)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2999..3587)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3079..3098)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3758..4618)
/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 4381..4400
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4619..4723)
/gene="bla"
/label=AmpR promoter
primer_bind 4791..4809
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
121 ttaaagagga gaaattaact atgggatcca gtcttcgcat gcatgaagac gcggccgcct
181 aggacccagc tttcttgtac aaagtggtaa gcttaattag ctgagcttgg actcctgttg
241 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
301 cgggcgtttt ttattggtga gaatccaagt aacaacacca tttaaatgga gtggttacaa
361 atggagtggt taattaaggc tagcttggcg agattttcag gagctaagga agctaaaatg
421 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
481 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
541 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
601 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
661 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
721 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
781 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
841 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
901 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
961 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
1021 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
1081 ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca
1141 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
1201 ggtgaacgct ctcctgagta ggacaaatcc gccctctaga ttacgtgcag tcgatgataa
1261 gctgtcaaac atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
1321 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
1381 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
1441 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
1501 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
1561 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
1621 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
1681 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
1741 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
1801 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
1861 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
1921 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
1981 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
2041 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
2101 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
2161 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
2221 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
2281 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
2341 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
2401 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
2461 tttgcaccat tcgatggtgt cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc
2521 atgatctaga gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
2581 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
2641 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
2701 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
2761 acatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
2821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
2881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
2941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
3001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
3061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
3121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
3241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
3301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
3361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
3421 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
3481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
3541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
3601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
3661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
3721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
3781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
3841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
3901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
3961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
4021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
4081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
4141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
4201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
4261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
4321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
4381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
4441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
4501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
4561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
4621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
4681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
4741 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
4801 tcacgaggcc ctttcgtctt cac
//
