質(zhì)粒序列
LOCUS Exported 6649 bp ds-DNA circular SYN 11-08-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6649)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-11 from SnapGene Viewer 2.8.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6649
/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..2628
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/note="MBP"
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
CDS 2677..2694
/codon_start=1
/product="recognition and cleavage site for Genenase(TM) I,
a subtilisin BPN' variant engineered for increased
substrate specificity (Carter and Wells, 1987)"
/note="Genenase(TM) I site"
/translation="PGAAHY"
primer_bind complement(2739..2755)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
terminator 3104..3190
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3282..3309
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 3329..3420
/gene="bla"
/note="AmpR promoter"
CDS 3421..4281
/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(4323..4836)
/direction=LEFT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 4947..5535
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 5721..5863
/note="bom"
/note="basis of mobility region from pBR322"
CDS complement(5965..6156)
/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 aaaactgaag aaggtaaact ggtaatctgg
1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
2461 ttggcgaaag atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg
2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg
2641 aacaacaaca acaataacaa taacaacaac ctcgggccgg gtgcggcaca ctacgtagaa
2701 ttcggatcct ctagagtcga cctgcaggca agcttggcac tggccgtcgt tttacaacgt
2761 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
2821 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
2881 ctgaatggcg aatggcagct tggctgtttt ggcggatgag ataagatttt cagcctgata
2941 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
3001 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
3061 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
3121 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
3181 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
3241 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
3301 tggccttttt gcgtttctac aaactctttt tgtttatttt tctaaataca ttcaaatatg
3361 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
3421 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
3481 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
3541 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
3601 gaagaacgtt ctccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
3661 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
3721 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
3781 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
3841 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
3901 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
3961 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
4021 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
4081 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
4141 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
4201 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
4261 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
4321 ttaccccggt tgataatcag aaaagcccca aaaacaggaa gattgtataa gcaaatattt
4381 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
4441 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
4501 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
4561 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca
4621 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
4681 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
4741 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
4801 cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaaa ggatctaggt gaagatcctt
4861 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
4921 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
4981 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
5041 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
5101 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
5161 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
5221 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
5281 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
5341 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
5401 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
5461 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
5521 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
5581 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
5641 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
5701 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
5761 tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
5821 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
5881 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
5941 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
6001 gcagctgcgg taaagctcat cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc
6061 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
6121 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
6181 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
6241 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
6301 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
6361 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
6421 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
6481 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
6541 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
6601 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgaaatt
//