質(zhì)粒序列
LOCUS Exported 6721 bp ds-DNA circular SYN 14-08-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMal-p2X
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6721)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-14 from SnapGene Viewer 2.8.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6721
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 3..80
/gene="lacI (mutant)"
/note="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 81..1163
/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"
misc_feature 1406..1434
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1442..1458
/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)."
CDS 1528..2703
/codon_start=1
/gene="malE"
/product="maltose binding protein from E. coli"
/note="MBP"
/note="This gene encodes a signal sequence that can direct
MBP to the bacterial periplasm."
/translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKD
PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
CDS 2752..2763
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
primer_bind complement(2811..2827)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
terminator 3176..3262
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3354..3381
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3401..3492
/gene="bla"
/note="AmpR promoter"
CDS 3493..4353
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(4395..4908)
/direction=LEFT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 5019..5607
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 5793..5935
/note="bom"
/note="basis of mobility region from pBR322"
CDS complement(6037..6228)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga
1501 gcacttcacc aacaaggacc atagcatatg aaaataaaaa caggtgcacg catcctcgca
1561 ttatccgcat taacgacgat gatgttttcc gcctcggctc tcgccaaaat cgaagaaggt
1621 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag
1681 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag
1741 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac
1801 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc
1861 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct
1921 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg
1981 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc
2041 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg
2101 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac
2161 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat
2221 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc
2281 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg
2341 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt
2401 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg
2461 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag
2521 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccactatgga aaacgcccag
2581 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact
2641 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag
2701 actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg gatcgaggga
2761 aggatttcag aattcggatc ctctagagtc gacctgcagg caagcttggc actggccgtc
2821 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
2881 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
2941 cagttgcgca gcctgaatgg cgaatggcag cttggctgtt ttggcggatg agataagatt
3001 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
3061 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
3121 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
3181 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
3241 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
3301 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
3361 catcctgacg gatggccttt ttgcgtttct acaaactctt tttgtttatt tttctaaata
3421 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
3481 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
3541 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
3601 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
3661 agttttcgcc ccgaagaacg ttctccaatg atgagcactt ttaaagttct gctatgtggc
3721 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
3781 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
3841 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
3901 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
3961 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
4021 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
4081 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
4141 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
4201 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
4261 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
4321 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
4381 ctttagattg atttaccccg gttgataatc agaaaagccc caaaaacagg aagattgtat
4441 aagcaaatat ttaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
4501 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
4561 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
4621 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
4681 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
4741 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
4801 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
4861 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtaa aaggatctag
4921 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
4981 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
5041 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
5101 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
5161 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
5221 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
5281 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
5341 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
5401 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
5461 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
5521 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
5581 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
5641 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
5701 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
5761 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
5821 ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg
5881 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
5941 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
6001 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
6061 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgcagcg attcacagat
6121 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
6181 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
6241 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
6301 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
6361 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
6421 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
6481 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
6541 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
6601 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
6661 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgaaat
6721 t
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