質(zhì)粒序列
LOCUS Exported 5440 bp ds-DNA circular SYN 31-7-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5440)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-7-31日 MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5440
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 244..266
/organism="Enterobacteria phage T7"
/mol_type="genomic DNA"
/db_xref="taxon:10760"
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"
CDS complement(205..237)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
RBS 244..266
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 281..305
/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(306..324)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 633..710
/gene="lacI"
/note="lacI promoter"
CDS 711..1793
/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 2602..2793
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2895..3037
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3223..3811)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3982..4842)
/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(4843..4947)
/gene="bla"
/note="AmpR promoter"
rep_origin complement(4974..5429)
/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 tccgacccat ttgctgtcca ccagtcatgc tagccatggt
241 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
301 attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc tctacgccgg
361 acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga
421 catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
481 gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc
541 attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca
601 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
661 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
721 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
781 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
841 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
901 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
961 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
1021 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
1081 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
1141 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
1201 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
1261 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
1321 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
1381 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
1441 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
1501 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
1561 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
1621 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
1681 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
1741 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
1801 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
1861 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
1921 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt
1981 ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc
2041 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc
2101 gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat cgtgctcctg
2161 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
2221 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
2281 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc
2341 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
2401 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
2461 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
2521 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat
2581 cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc
2641 tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa
2701 caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc
2761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
2821 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
2881 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
2941 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat
3001 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
3061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
3121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
3181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
3241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
3301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
3361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
3421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
3481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
3541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
3601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
3661 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
3721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
3781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
3841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
3901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
3961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
4021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
4081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
4141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
4201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
4261 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
4321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
4381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
4441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
4501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
4561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
4621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
4681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
4741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
4801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
4861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
4921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt
4981 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
5041 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
5101 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
5161 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
5221 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
5281 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
5341 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
5401 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//