LOCUS Exported 6624 bp ds-DNA circular SYN 28-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pRH2521
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6624)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 28, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6624
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 63..182
/label=Pimyc promoter
terminator 197..224
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
misc_feature 244..423
/label=Pmyc1 promoter
protein_bind 424..441
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_RNA 503..578
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS complement(738..1361)
/codon_start=1
/label=TetR
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
misc_feature complement(1389..1508)
/label=Pimyc promoter
terminator 1588..1615
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
rep_origin complement(1876..2464)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2843..4006)
/codon_start=1
/label=Hyg
/translation="MPGPFTPAAAERPPPGPHRRRPARSGLAAAGFGAGPGLPSAGGAP
GGRPPGAGGGAGRPGASGAGGGVRRPPEELRQFLRIGEAGEIQRGLLEHLEVVQEGEGE
QFAGEVLGPLPLRPVEQRGQDLAVAPEGVEMQLHQAVAGVSRIDVGEVDDPGDLGRGQV
HEDVGPVQVAVDEPGFAAGQQRVHVRQPVLQAVQQPGREVAPPAVVLDGRRAAFPQQFR
EDLGMGGEHGVPGQRHPVQPAEHPAEFAGQGEQRVPVGRAVHRGPPGGAGHPAHHQVGP
RPGSGAGPQLAAAEEAGHRHRGVRQDRVRLRLRREALRTAPVLAEQLDHRAGLADQYGV
GALAGHPQHRRHRQPELLQGSAGQRLPEFLVVPQARVGIIRINTVEK"
misc_feature complement(4090..6616)
/label=oriM
ORIGIN
1 aacgttctcg gctcgatgat ccccgggggc cccccctcga ggtcgacggt atcgataata
61 ttggatcgtc gcaccgggtt aagcctggag tgcggtggtg cctggtcggc attttcgcag
121 tcgagggctc tcgtgtagcc tgggcgagtt gccgacgcag gcgaccctcc tgccacggat
181 ccgtgattag ctaagcagaa ggctatcctg acggatggcc tttttgcgtt taatactgtt
241 taaactctag aaatattgga tcgtcggcac cgtcacggcc gtgggaggcg gcacgatccg
301 cgacgtgatg atcggccgca tccccacggt gctgcgcagt gagctctacg ccatcccggc
361 gttgatctgt gcgttcgcac gcacaggccc ggtgtgagaa gggtctctga cgagcgggag
421 aactccctat cagtgataga gtttgtcctc cctatcagtg atagataggc tatgggagtg
481 tcttcgagac agaggaagac atgttttaga gctagaaata gcaagttaaa ataaggctag
541 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttatcgat ttaattaata
601 tgcatcaatt gatttaaata ctagtgttaa ctatttaatt ggggacccta gaggtcccct
661 tttttatttt aaaaattttt tcaaggggac ctctagggtc cccagctggc tagcgagtca
721 tgaggtcacc atggtcatta ggagccgctc tcgcacttca gctgcttctc caagccgcag
781 atgatcagct ccaggccgaa caagaaggcc ggctcggcgc cctggtggtc gaacagctcg
841 atggcctggc gcagcaacgg cggcatgctg tcggtggtcg gggtctcgcg ctcctccttg
901 gcgacctggt gctcctggtc ctccaacacg cagcccaggg tgaagtggcc cacggccgac
961 agggcgtaca acgcgttctc cagcgagaag ccctgctggc acaggaaggc caactggttc
1021 tccagggtct cgtactgctt ctcggtcggc cgggtgccca ggtgcacctt ggcgccgtcg
1081 cggtggctca gcaaggcgca gcggaagctc ttggcgttgt tgcgcaggaa gtcctgccag
1141 ctctcgccct ccagcgggca gaagtgggtg tggtggcggt ccaacatctc gatggccagg
1201 gcgtcgagca gggcccgctt gttcttgacg tgccagtaca gggtcggctg ctcgacgccc
1261 agcttctggg ccagcttgcg ggtggtcagg ccctcgatgc cgacctcgtt cagcagctcc
1321 agggcggagt tgatgacctt ggacttgtcc aggcgggaca ttgcgaagtg attcctcctg
1381 atggtaccgg atccgtggca ggagggtcgc ctgcgtcggc aactcgccca ggctacacga
1441 gagccctcga ctgcgaaaat gccgaccagg caccaccgca ctccaggctt aacccggtgc
1501 gacgatccaa tattacgaga tctgatatcc tcgaggcatg ccacgtgcct gcaggtttaa
1561 acgcggccgc aaacagtatt aaacgcaaaa aggccatccg tcaggatggc cttctgctta
1621 gctaatcaac tagttctagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
1681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
1741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
1801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
1861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
1921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
1981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
2041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
2101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
2161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
2221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
2281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
2341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
2461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
2521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
2641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
2701 ctgactcccc gacgtggccg accagcccgt catcgtcaac gcctgatccg cggtgcggac
2761 aggccgtgtc gcgaccggcc gtgcggaatt aagccggccc gtaccctgtg aatagaggtc
2821 cgctgtgaca caagaatccc tgttacttct cgaccgtatt gattcggatg attcctacgc
2881 gagcctgcgg aacgaccagg aattctggga gccgctggcc cgccgagccc tggaggagct
2941 cgggctgccg gtgccgccgg tgctgcgggt gcccggcgag agcaccaacc ccgtactggt
3001 cggcgagccc ggcccggtga tcaagctgtt cggcgagcac tggtgcggtc cggagagcct
3061 cgcgtcggag tcggaggcgt acgcggtcct ggcggacgcc ccggtgccgg tgccccgcct
3121 cctcggccgc ggcgagctgc ggcccggcac cggagcctgg ccgtggccct acctggtgat
3181 gagccggatg accggcacca cctggcggtc cgcgatggac ggcacgaccg accggaacgc
3241 gctgctcgcc ctggcccgcg aactcggccg ggtgctcggc cggctgcaca gggtgccgct
3301 gaccgggaac accgtgctca ccacccattc cgagatcttc ccggaactgc tgcgggaacg
3361 ccgcgcggcg accgtcgagg accaccgcgg gtggggctac ctctcgcccc ggctgctgga
3421 ccgcctggag gactggctgc cggacgtgga cacgctgctg gccggccgcg aaccccggtt
3481 cgtccacggc gacctgcacg ggaccaacat cttcgtggac ctggccgcga ccgaggtcac
3541 cgggatcgtc gacttcaccg acgtctatgc gggagactcc cgctacagcc tggtgcaact
3601 gcatctcaac gccttccggg gcgaccgcga gatcctggcc gcgctgctcg acggggcgca
3661 gtggaagcgg accgaggact tcgcccgcga actgctcgcc ttcaccttcc tgcacgactt
3721 cgaggtgttc gaggagaccc cgctggatct ctccggcttc accgatccgg aggaactggc
3781 gcagttcctc tgggggccgc cggacaccgc ccccggcgcc tgacgccccg ggccgcccgg
3841 cgccgccccc ggcccccggc ggccggccgc ccggagcccc gcccgcgctc gggagccccg
3901 ggcccgcgcc gaagcccgct gctgcgagcc cggagcgggc cggccgacgg cggtgcgggc
3961 ccggcggcgg acgctcagca gcggcgggcg tgaaaggccc tggcatcctc gatcatctcc
4021 tccagggtgg tcggccgagc ttccatccca gctcggcaag gatcaacgag gacagtcgca
4081 cgacgaagtt cttctggatc gcgcccgtgc tggaagcact caacctcgaa gcgtgtggtt
4141 gcggagccat ctagcaacca cacgaaacat gcgcaacgaa ccgcgcaacg aacaacgcct
4201 agaactggcc ctagatgagc tgactcgtat cgttggtaaa cctagtttga ccagcatgtt
4261 ttaactacgt tcggtgagct gtcaacgggg cctgtaacgg cacaacgaac cgtgcaacga
4321 gagtggccac ggatgccacc acaggcacta caacggagtt cgccacgtac atcaccacaa
4381 ccaccgattc tggcggtgag ctccccgata ttcagcggaa atggcttggt atcgaccaag
4441 attcgtagaa ccccgtctcg tctggctggt attcaaaacg ggcgcaacga aacacgcaac
4501 gagacaggca tggcccaaac cagaaaacta gcgtctacca ggacttttac ctgtccgacc
4561 cgttgcaacg gaacccccca cggaaccccc gcgacacccg ctccccaatt gcgttagaac
4621 agcggtggat tgtcggcttc gttgtgggcc ttttgagccg cttcctgttc tgccgcacgc
4681 tctttcctcg cccgatagcc gagtcgctta acggtgtcca gatgcagccc gaaatgtttg
4741 gccgtttgcg gccaagagtg gccctcgtcg tcgtgatagg cgcggatgcg ttcgcggcgt
4801 gcagcctgct cggcgagcca ctcgctgcgt tcctgcgcca cgagccggac gacgtggcgt
4861 tcggatagtc cggtgattcg agcgccttcg gcggcggtca cgcgccgctt tttgcggaca
4921 gtcggctgcc ggttgtagcc gtcgctgtag ccgtcgctca tagcaatgcc tccatggctg
4981 acgcggactt tgcgcgccgc gcaactgtgc tcgccgccgt gcgcgctgct gcgcccttcc
5041 gcgagatggc cgactggcgc gcactgagtg tggcctcgta gaccacgatc ccgtccgccc
5101 aaatgcgcga cttggttgtg atccaacgcc aaatgctgtt ggcgatggcg cggacctcgc
5161 tgtccggtag cggtccggga cacacgtcgt tgcacgggaa ttcggcgttt cgcgcgtggc
5221 actcggcata gatcgcgcgg ccgagtccgt ccacgttccg ggtcggcagg tagatccgca
5281 tgagggcggg acgataggcc cacaacctga cggaatcgaa cagtgcgcaa ttccgcccta
5341 gcggcgtcgg agccgctttg tacgtggtct gctgacgcca gcgcggcggt ggcatgttcg
5401 cgccgagctc ggcctcgatg tggctgagtg tgtagagatc tgagtggagc cattccgttt
5461 cccaggcgat gtggccgggg tttttggtca tgaggcctga gtaactgcgg tcgccgtcga
5521 cggcgcgccg aaggccttcg gcgcacgccg ccatgtatgc gagcggctta cgccgcgcgt
5581 attcggtgcg tggaacaggg gcgttgagtg cccacactgc gtgtgcgtgg ccgttggcgc
5641 gattgcccac gatcgcgttg ggcagcggat gggacccccg ggcgctgagc gctcggagcg
5701 ctgcgtctgg atggtctacg tccacgacca gcaggtttgc cagcgctgtt gggttcgcct
5761 cgatgtaccg gcggcctagg gccgacgcgc ggctttggcg gtagatcccc tcgagcagat
5821 cgtcgcttgc cagcggccag tacggcagcc agagctgctc aaattcgtcg gcgacgtggc
5881 tcacgcttgg tagtagacca cgattaatca ccggtgtatg gtccgacacg agctccaagt
5941 cagatatttc gctgaggggc caccccacaa ctgcacactc ccccgctctc ccgtcgagcc
6001 ctggtggtgg aacaccagcg acagccgagc acccccaacc acctgtacca accaggagga
6061 acacatgcgt cgtttcgagg acgtttccgg gccgctgaga gccgctgtgg cggccgtaca
6121 cgccgcctta gacccgttag accccctgcc gcctgaatgc gcgggtacga gccacacagc
6181 gcccgaactt acggagctgg tgggctcacc tggctttatg gcgtacgaat cggctgtgtg
6241 cgacctgttg ggcgaggtga ggtacgcgct actcacgctg gcaagggcga cagagccgcc
6301 ccaccgagcc cgcacggccg cgcgcggtgt caacaaccgg gtgagtcgtg cacaccagca
6361 ggtgttcgag gcttggctcg aagtgcagga catcgtggcg aacgccgccc gatgagccgc
6421 gccttacgct ggctgccagc cgttcgcggg ctggttggtg cagcgcgtcg agcggttaga
6481 ggccctgcgg tgttccacca ccgcagggcc tcgccctttt taaggctgaa tttgcttgtc
6541 tccgaatcca actggcttgt ccaagggtgt atctacgctt aatccaaagt tcaaacgagg
6601 ggattacaca tgaccaactt cgat
//