質(zhì)粒序列
LOCUS Exported 5307 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET-24d
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5307)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5307
/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"
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"
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"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
rep_origin complement(3223..3811)
/direction=LEFT
/note="ori"
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS 3933..4748
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(4841..5296)
/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 gtcatgaaca ataaaactgt
3901 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
3961 gctctaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
4021 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
4081 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
4141 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
4201 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
4261 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
4321 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
4381 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
4441 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
4501 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
4561 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
4621 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
4681 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
4741 ttttctaaga attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa
4801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt
4861 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
4921 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
4981 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
5041 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
5101 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
5161 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
5221 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
5281 ccgctacagg gcgcgtccca ttcgcca
//