質(zhì)粒序列
LOCUS Exported 7309 bp ds-DNA circular SYN 06-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET-44c
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7309)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7309
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 260..346
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(512..529)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(536..568)
/codon_start=1
/product="HSV (herpes simplex virus) epitope tag"
/note="HSV tag"
/translation="QPELAPEDPED"
CDS complement(679..693)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(712..729)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(745..789)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(799..816)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(832..2316)
/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"
CDS complement(2323..2340)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
protein_bind 2397..2421
/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(2422..2440)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2753..2830
/gene="lacI"
/note="lacI promoter"
CDS 2831..3913
/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 4485..4676
/codon_start=1
/gene="rop"
/product="Rop protein"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(5106..5694)
/direction=LEFT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
promoter 5775..5867
/gene="bla"
/note="AmpR promoter"
CDS 5868..6728
/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(6843..7298)
/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 tcagcggata catatttgaa tgtatttaga aaaataaaca
121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
661 gatccacgag ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg
721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc
781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc
841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa
901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac
961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc
1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc
1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa
1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag
1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg
1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc
1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt
1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac
1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat
1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc
1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg
1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac
1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc
1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa
1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc
1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg
1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac
1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac
2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt
2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct
2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt
2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga
2281 tacggcttca actacagcca aaatttcttt gttcatacta gaatggtgat ggtggtgatg
2341 gctgctgccc atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
2401 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg
2461 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
2521 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
2581 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
2641 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
2701 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
2761 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
2821 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
2881 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
2941 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
3001 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
3061 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
3121 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
3181 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
3241 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
3301 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
3361 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
3421 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
3481 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
3541 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
3601 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga
3661 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
3721 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
3781 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
3841 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
3901 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
3961 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
4021 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
4081 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
4141 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
4201 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
4261 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
4321 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
4381 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
4441 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
4501 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
4561 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
4621 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
4681 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
4741 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
4801 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
4861 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
4921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
4981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
5041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
5101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
5161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
5221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
5281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
5341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
5401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
5461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
5521 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
5581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
5641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
5701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
5761 ttggtcatga cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
5821 acaataaccc tgataaatgc ttcaatcatg attgaaaaag gaagagtatg agtattcaac
5881 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
5941 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
6001 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
6061 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
6121 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
6181 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
6241 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
6301 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
6361 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
6421 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
6481 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
6541 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
6601 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
6661 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
6721 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
6781 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgcca tcgtgccgcc
6841 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
6901 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
6961 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
7021 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
7081 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
7141 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
7201 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
7261 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
//