质粒序列
LOCUS Exported 6190 bp ds-DNA circular SYN 10-8-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6190)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-10 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6190
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 1123..1145
/organism="Enterobacteria phage T7"
/mol_type="genomic DNA"
/db_xref="taxon:10760"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(150..173)
/codon_start=1
/product="8xHis affinity tag"
/note="8xHis"
/translation="HHHHHHHH"
CDS complement(252..266)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(282..326)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(342..359)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(369..386)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(408..1115)
/codon_start=1
/gene="dsbC"
/product="bacterial periplasmic protein disulfide
isomerase"
/note="DsbC"
/translation="MKKGFMLFTLLAAFSGFAQADDAAIQQTLAKMGIKSSDIQPAPVA
GMKTVLTNSGVLYITDDGKHIIQGPMYDVSGTAPVNVTNKMLLKQLNALEKEMIVYKAP
QEKHVITVFTDITCGYCHKLHEQMADYNALGITVRYLAFPRQGLDSDAEKEMKAIWCAK
DKNKAFDDVMAGKSVAPASCDVDIADHYALGVQLGVSGTPAVVLSNGTLVPGYQPPKEM
KEFLDEHQKMTSGK"
RBS 1123..1145
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 1160..1184
/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)."
promoter complement(1185..1203)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1516..1593
/gene="lacI"
/note="lacI promoter"
CDS 1594..2676
/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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS 3485..3676
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3778..3920
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4106..4694)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4816..5631
/codon_start=1
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(5724..6179)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
181 gcggccgcaa gcttgtcgac ggagctcgaa ttcggatccg atatcgccat ggttgaggag
241 aagcccgggc tcttgtcgtc gtcatcggta cccagatctg ggctgtccat gtgctggcgt
301 tcgaatttag cagcagcggt ttctttcata ccaattgcag tactaccgcg tggcaccaga
361 cccgcggagt gatggtgatg gtgatgacca gaaccactag ttgatccttt accgctggtc
421 attttttggt gttcgtcgag aaattctttc atctctttcg gcggctggta acccggaaca
481 agtgtgccat tgctcagcac aactgccgga gtaccgctaa cgccaagctg gacgccaagt
541 gcgtaatggt cggcaatatc cacgtcgcaa ctggctggtg cgacgctttt acctgccatc
601 acatcatcaa acgctttgtt tttatctttc gcacaccaga tagctttcat ttctttctct
661 gcatcgctgt ccagcccctg gcgcgggaaa gcaagataac gcacggtgat ccccagcgcg
721 ttgtagtctg ccatttgctc atgcagtttg tggcagtaac cacaggtaat atcagtaaac
781 acggtgatga cgtgtttttc ctgcggcgct ttataaacga tcatctcttt ttcaagcgca
841 ttcaactgct ttaacagcat cttattggtg acattgaccg gagccgtgcc actaacgtca
901 tacattggcc cctgaatgat atgtttacca tcatcggtga tgtacaacac gccgctgtta
961 gtcagaactg tcttcatgcc agctacaggc gcgggctgaa tatcgctgct tttgatgccc
1021 attttggcta acgtttgttg aattgccgcg tcatcagcct gagcaaagcc tgaaaacgcc
1081 gctaacaaag taaacaacat aaaacctttc ttcatatgta tatctccttc ttaaagttaa
1141 acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta
1201 ttaatttcgc gggatcgaga tcgatctcga tcctctacgc cggacgcatc gtggccggca
1261 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
1321 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
1381 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
1441 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
1501 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
1561 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
1621 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
1681 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
1741 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
1801 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
1861 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
1921 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
1981 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
2041 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
2101 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
2161 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
2221 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
2281 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
2341 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggacatctcg
2401 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
2461 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
2521 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
2581 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
2641 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
2701 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
2761 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
2821 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
2881 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
2941 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
3001 cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
3061 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
3121 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
3181 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
3241 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
3301 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
3361 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
3421 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
3481 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
3541 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
3601 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
3661 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
3721 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
3781 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
3841 gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa
3901 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
3961 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
4021 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
4081 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
4141 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
4201 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
4261 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
4321 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
4381 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
4441 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
4501 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
4561 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
4621 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
4681 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
4741 aaaactcacg ttaagggatt ttggtcatga acaataaaac tgtctgctta cataaacagt
4801 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cttgctctag gccgcgatta
4861 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
4921 tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa
4981 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
5041 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
5101 ttactcacca ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat
5161 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
5221 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
5281 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
5341 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
5401 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
5461 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
5521 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
5581 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta agaattaatt
5641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
5701 atttccccga aaagtgccac ctgaaattgt aaacgttaat attttgttaa aattcgcgtt
5761 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
5821 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
5881 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
5941 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
6001 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
6061 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
6121 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
6181 ccattcgcca
//