| 启动子: | J23100 promoter，PcpcG2-172 |
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| 复制子: | ori |
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| 终止子: | rrnB T1 terminator，T7Te terminator |
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| 质粒分类: | 基因文库质粒;其它基因质粒;其它种源质粒 |
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| 质粒大小: | 3936bp |
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| 原核抗性: | 氯霉素Chl（50μg/ml） |
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| 筛选标记: | superfolder GFP |
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| 克隆菌株: | DH5α等大肠杆菌 |
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| 培养条件: | 37℃，有氧，LB |
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Expresses CcaR constitutively and sfGFP under the PcpcG2-172 promoter.
LOCUS Exported 3936 bp ds-DNA circular SYN 15-AUG-2017
DEFINITION synthetic circular DNA
KEYWORDS pSR58.6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3936)
TITLE Direct Submission
JOURNAL Exported Aug 15, 2017 from Plasmid
FEATURES Location/Qualifiers
source 1..3936
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..35
/label=J23100 promoter
/note="Constitutive promoter J23100"
misc_feature 36..69
/label=RBS
/note="sRBS (250)"
CDS 70..774
/codon_start=1
/label=ccaR
/translation="MRILLVEDDLPLAETLAEALSDQLYTVDIATDASLAWDYASRLEY
DLVILDVMLPELDGITLCQKWRSHSYLMPILMMTARDTINDKITGLDAGADDYVVKPVD
LGELFARVRALLRRGCATCQPVLEWGPIRLDPSTYEVSYDNEVLSLTRKEYSILELLLR
NGRRVLSRSMIIDSIWKLESPPEEDTVKVHVRSLRQKLKSAGLSADAIETVHGIGYRLA
NLTEKSLCQGKN"
terminator 795..866
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 882..909
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_feature 1033..1204
/label=PcpcG2-172
RBS 1205..1216
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1223..1936
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
terminator 1963..2034
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2050..2077
/label=T7Te terminator
/note="phage T7 early transcription terminator"
rep_origin complement(2279..2867)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator 2955..3049
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(3070..3729)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(3730..3832)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
ORIGIN
1 ttgacggcta gctcagtcct aggtacagtg ctagctgtta aacgctcctc gtaagaacac
61 gggaaagcaa tgagaattct tttagtggag gatgatttgc cgctggcgga aacccttgct
121 gaagcattga gtgaccagct ttacaccgtt gatattgcca ccgacgcttc cctcgcctgg
181 gactatgcct cccgactgga atatgacctc gttattttgg atgtgatgct gccggagttg
241 gacgggatta ccctctgtca aaaatggcga tcgcacagtt atttaatgcc aattttgatg
301 atgacagcca gggatacgat caatgataaa atcacgggct tggatgcggg ggcggatgat
361 tatgtggtca agccagtgga tttgggggag ttatttgcca gggtgcgagc tttgttgcgt
421 cggggttgtg caacgtgcca accagtttta gagtgggggc caatcaggtt ggatccaagc
481 acctatgaag ttagttatga caatgaggtt ttgtctttga cccgcaagga atacagcatt
541 ctggaattac tactccgcaa tggccgtcgg gtgctaagtc ggagcatgat tatcgatagt
601 atctggaagt tggagagtcc cccagaggaa gatacggtta aggtgcatgt gcggagtttg
661 cgacaaaaat taaaaagtgc cggtttatca gcagatgcca ttgaaacggt ccatggcatt
721 gggtatcgtc tggccaattt aacggaaaaa tctttgtgcc aagggaaaaa ctagtaataa
781 tctagaccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
841 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
901 ctttctgcgt ttatagtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc
961 tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg
1021 gttagctagt caagcccatt gtgcttttct ctatcaacct cagcttacct gaaggggtga
1081 acaggtctgg gttaattcat gttgcgaaat gtaacagttt tagtcgcatc agctaacttt
1141 ccgatttctt tacgattttc tccccctttt cttcaatttt actttgttag gatcgcattt
1201 ttaaaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc
1261 tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg
1321 tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta ctggtaaact
1381 gccggtacct tggccgactc tggtaacgac gctgacttat ggtgttcagt gctttgctcg
1441 ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg aaggctatgt
1501 gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa
1561 atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact ttaaagaaga
1621 cggcaatatc ctgggccata agctggaata caattttaac agccacaatg tttacatcac
1681 cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca acgtggagga
1741 tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg atggtcctgt
1801 tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag atccgaacga
1861 gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca cgcatggtat
1921 ggatgaactg tacaaatgat gataataatc tagaccaggc atcaaataaa acgaaaggct
1981 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
2041 agtcacactg gctcaccttc gggtgggcct ttctgcgttt ataggtaccc gtggatcctc
2101 tagagtcgac ctgcaggcat gcaagcttcc taggcgttcg gctgcggcga gcggtatcag
2161 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
2221 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
2281 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
2341 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
2401 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
2461 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
2521 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
2581 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
2641 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
2701 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
2761 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
2821 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
2881 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
2941 tgactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
3001 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
3061 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
3121 tctgccgaca tggaagccat cacagacggc atgatgaacc tgaatcgcca gcggcatcag
3181 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
3241 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
3301 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
3361 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
3421 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
3481 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc
3541 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
3601 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
3661 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
3721 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
3781 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
3841 ttttcgccag atatcgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
3901 aggcgtatca cgaggccctt tcgtcttcac ctcgag
