LOCUS Exported 6001 bp ds-DNA circular SYN 28-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6001)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 28, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6001
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 141..159
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 160..184
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
RBS 199..221
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 240..257
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 276..995
/codon_start=1
/label=VENUS
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
terminator 1086..1133
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1170..1625
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(1718..2533)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 2655..3243
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 3429..3571
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(3673..3864)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(4673..5755)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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"
promoter complement(5756..5833)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
61 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
121 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
181 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggcagcagcc
241 atcatcatca tcatcacagc agcggccgcg gatccatggt gtccaagggc gaggagctgt
301 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
361 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct
421 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc
481 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
541 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
601 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
661 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
721 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
781 gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
841 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga
901 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
961 ggatcactct cggcatggac gagctgtaca agtgactcga gcaccaccac caccaccact
1021 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
1081 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
1141 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
1201 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
1261 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
1321 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
1381 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
1441 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
1501 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
1561 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
1621 aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
1681 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
1741 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
1801 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
1861 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
1921 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
1981 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
2041 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
2101 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
2161 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
2221 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
2281 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
2341 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
2401 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
2461 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
2521 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
2581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
2641 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
2701 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
2761 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
2821 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
2881 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
2941 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
3001 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
3061 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
3121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
3181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
3241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
3301 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
3361 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
3421 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
3481 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
3541 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
3601 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
3661 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
3721 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
3781 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
3841 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
3901 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
3961 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
4021 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
4081 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
4141 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
4201 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
4261 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
4321 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
4381 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
4441 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
4501 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
4561 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
4621 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
4681 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
4741 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
4801 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
4861 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
4921 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
4981 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
5041 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
5101 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
5161 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
5221 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
5281 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
5341 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
5401 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
5461 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
5521 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
5581 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
5641 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
5701 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
5761 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
5821 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
5881 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
5941 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
6001 c
//