质粒序列
LOCUS Exported 5369 bp ds-DNA circular SYN 26-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET-21
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Friday, August 26, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5369
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
protein_bind 210..234
/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(235..253)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 562..639
/gene="lacI"
/note="lacI promoter"
CDS 640..1722
/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 2531..2722
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2824..2966
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3152..3740)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3911..4771)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4772..4876)
/gene="bla"
/note="AmpR promoter"
rep_origin complement(4903..5358)
/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 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tcctagaggg gaattgttat ccgctcacaa ttcccctata
241 gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg
301 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
361 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
421 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg
481 cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata
541 agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca
601 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
661 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
721 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
781 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
841 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
901 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
961 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
1021 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
1081 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
1141 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
1201 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
1261 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
1321 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
1381 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
1441 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
1501 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
1561 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
1621 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
1681 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
1741 aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc
1801 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
1861 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
1921 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
1981 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
2041 cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt cgttgaggac
2101 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg
2161 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt
2221 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt
2281 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
2341 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag
2401 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt
2461 gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac
2521 ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag
2581 ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat
2641 ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg
2701 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
2761 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
2821 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
2881 gcatcagagc agattgtact gagagtgcac catatatgcg gtgtgaaata ccgcacagat
2941 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
3001 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
3061 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
3121 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
3181 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
3241 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
3301 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
3361 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
3421 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
3481 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
3541 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
3601 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
3661 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
3721 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
3781 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
3841 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
3901 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
3961 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
4021 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
4081 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
4141 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
4201 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
4261 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
4321 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
4381 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
4441 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
4501 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
4561 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
4621 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
4681 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
4741 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
4801 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
4861 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
4921 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
4981 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
5041 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
5101 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
5161 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
5221 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
5281 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
5341 cgccgctaca gggcgcgtcc cattcgcca
//