质粒序列
LOCUS Exported 7275 bp ds-DNA circular SYN 06-SEP-2016
DEFINITION synthetic circular DNA
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7275)
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7275
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 25..72
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 140..167
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 259..345
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(511..528)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(535..567)
/codon_start=1
/product="HSV (herpes simplex virus) epitope tag"
/note="HSV tag"
/translation="QPELAPEDPED"
CDS complement(680..694)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(713..730)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(746..790)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(800..817)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(833..2317)
/codon_start=1
/gene="nusA"
/product="transcription elongation protein (N utilization
substance protein A)"
/note="NusA"
/note="highly soluble in E. coli"
/translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
RBS 2325..2347
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 2362..2386
/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(2387..2405)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2718..2795
/gene="lacI"
/note="lacI promoter"
CDS 2796..3878
/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 4450..4641
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 4743..4885
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5071..5659)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 5740..5832
/gene="bla"
/note="AmpR promoter"
CDS 5833..6693
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(6808..7263)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 tccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag
61 gggttatgct agttattgct cagcggatac atatttgaat gtatttagaa aaataaacaa
121 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct taatttgatg
181 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt
241 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
301 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggcagttcc
361 ctactctcgc atggggagac cccacactac catcggcgct acggcgtttc acttctgagt
421 tcggcatggg gtcaggtggg accaccgcgc tactgccgcc aggcaaattc tgttttatca
481 gcctaggaac gcccaactta attaacatta gtggtggtgg tggtggtgct cgagatcctc
541 tgggtcttca ggagcgagtt ctggctggct tgcacgtgta tacagctgtg cggccgcaag
601 cttcgaacgc gtatcgatgg taccgtcgac gtcctgcagg cgcgcctgta cagaattcgg
661 atccacgagc tcccggactc ttgtcgtcgt catcaatcgt accagaaccc gcgctgcccc
721 gggggaccag accagttggc ggtggcgagt ccatgtgctg gcgttcaaat ttcgcagcag
781 cggtttcttt acccgcggag tgatggtgat ggtgatgacc agaaccacta gtcgcttcgt
841 caccgaacca gcaaatatta cgggcagcca taatcagtgc tccggctttt tcgtcggtca
901 acccttcgat atcagccaga tcatcaatgc cctgttcggc gagatcttcc agcgtacaaa
961 cgccacgggc ggccagtttg aatgccaaat cacgatctac cccttcaagg ttcagcagat
1021 cgtcagccgg tttgttatca ccgaggcttt cttcctgggc ctgtgcaatg gtggccagtg
1081 catttttagc acgctcgcgc agtgcttcaa cggtcggctc atcaaggcct tcgatttcca
1141 acagctcttt catcggcaca taggccaatt cttccagcgt cgagaagcct tcttctacca
1201 gaacagtcgc gaagtcttcg tcgatgtcga gatatttggt gaaggtgtcg atcgctgcgt
1261 gcgcttccgc ctgatgctta gcttgcaggt cgtcaacggt catcacgttg agttcccaac
1321 cgctcagttg cgaagccaga cgcacgttct gaccgttacg gccaatcgcc tgcgccagat
1381 taccggcttc aacggcgatg tccatggtgt gtttatcttc atccaccacg atagaagcaa
1441 cgtctgccgg tgccattgcg ttaatcacga actgcgccgg gttatcatcc cacaggacga
1501 tatcgatacg ctcgccaccc agttcagtag acaccgcctg aacacgcgcg ccacgcatac
1561 ctacgcaagc acctaccgga tcgatacgtt tatcgttggt tttcaccgcg attttcgcac
1621 gagaacccgg atcgcgagcc gctgctttaa tttcaatcac ttcttcgccg atttctggca
1681 cttcaatacg gaacagttcg atcagcattt ccggcttgga acgagtgacg aacagttgcg
1741 cgccacgcgc ttccgggcga acggaataga gcacgccacg aacgcggtcg ccagggcgga
1801 agttttcacg cggcagcata tcttcgcgca ggatcacggc ttcagcgttg ttgcccagat
1861 ccagagagat gttgtcgcgg tttacttttt tcaccacgcc ggtgatgatt tcaccttcgt
1921 gttcacggaa ctgatcaacc accatcgcac gttcggcttc acgcactttc tgcacgataa
1981 cctgttttgc cgtctgggta gtgatacggt caaaggtaac agactcaatc tgatcttcaa
2041 cgtaatcgcc caggttcagg ctttcatctt cataacgtgc ggcttcaagg gtgatttcct
2101 tggtcggctg ggtgacttca tcaacaacta accagcgacg gaaagtgtca aaatcaccgc
2161 ttttgcgatc gatctgtacg cggacgtcga tctcttgttc atattttttc tttgttgctg
2221 tcgccagcgc gctttccaat gcttcgaaaa tcttctcgcg aggtagcgcc ttttcattgg
2281 atacggcttc aactacagcc aaaatttctt tgttcatatg tatatctcct tcttaaagtt
2341 aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg
2401 tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca tcgtggccgg
2461 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
2521 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg
2581 ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc
2641 ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg
2701 agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat
2761 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
2821 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
2881 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
2941 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
3001 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
3061 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
3121 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
3181 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
3241 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
3301 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
3361 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
3421 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
3481 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
3541 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggacatct
3601 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
3661 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
3721 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
3781 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
3841 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt
3901 tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt caacccagtc
3961 agctccttcc ggtgggcgcg gggcatgact agcatgatcg tgctcctgtc gttgaggacc
4021 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
4081 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
4141 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
4201 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
4261 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
4321 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
4381 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
4441 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
4501 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
4561 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
4621 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
4681 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
4741 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
4801 catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac cgcacagatg
4861 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
4921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
4981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
5041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
5101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
5161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
5221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
5281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
5341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
5401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
5461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
5521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
5581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
5641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
5701 gaacgaaaac tcacgttaag ggattttggt catgacctat ttgtttattt ttctaaatac
5761 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa tcatgattga
5821 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
5881 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
5941 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
6001 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
6061 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
6121 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
6181 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
6241 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
6301 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
6361 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
6421 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
6481 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
6541 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
6601 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
6661 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
6721 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
6781 gataatctca tgccatcgtg ccgccagaaa ttgtaaacgt taatattttg ttaaaattcg
6841 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
6901 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
6961 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
7021 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
7081 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
7141 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
7201 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
7261 cgtcccattc gccaa
//