LOCUS Exported 9847 bp ds-DNA circular SYN 11-OCT-2017
DEFINITION synthetic circular DNA
KEYWORDS pCAMBIA35S-4×Myc-MCS-3×FLAG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9847)
TITLE Direct Submission
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9847
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 234..579
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 599..718
/codon_start=1
/product="3 tandem Myc epitope tags"
/label=4×Myc
/translation="EQKLISEEDLEQKLISEEDLEQKLISEEDLEQKLISEEDL"
misc_feature 719..802
/label=MCS
/note="pUC18/19 multiple cloning site"
CDS 803..868
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
terminator 894..1146
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 1168..1192
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 2492..3121
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 3555..4622
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 4688..4882
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 5226..5366
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(5552..6140)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6227..7021)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=Kan
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 7446..7470
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 7548..7722
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(7762..8787)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=Hyg
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(8855..9532)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 9723..9744
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9759..9789
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9797..9813
/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)."
ORIGIN
1 agcagcttga gcttggatca gattgtcgtt tcccgccttc agtttagctt catggagtca
61 aagattcaaa tagaggacct aacagaactc gccgtaaaga ctggcgaaca gttcatacag
121 agtctcttac gactcaatga caagaagaaa atcttcgtca acatggtgga gcacgacaca
181 cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc aattgagact
241 tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc tatctgtcac
301 tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca ttgcgataaa
361 ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg acccccaccc
421 acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga
481 tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct
541 tcctctatat aaggaagttc atttcatttg gagagaacac gggggactct tgaccatgga
601 acaaaaacta atatcagagg aagatttgga acagaaacta atcagtgaag aagatttaga
661 gcaaaagtta atttccgagg aggacttaga gcagaagttg atttcagaag aagatctggg
721 gcccataggc ctgaattcga gctcggtacc cggggatcct ctagagtcga cctgcaggca
781 tgcaagctta ctagtcacgc gtgactacaa agaccatgac ggtgattata aagatcatga
841 catcgattac aaggatgacg atgacaagta aggtgaccag ctcgaatttc cccgatcgtt
901 caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta
961 tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt
1021 tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag
1081 aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac
1141 tagatcggga attaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga
1201 aaagagcgtt tattagaata acggatattt aaaagggcgt gaaaaggttt atccgttcgt
1261 ccatttgtat gtgcatgcca accacagggt tcccctcggg atcaaagtac tttgatccaa
1321 cccctccgct gctatagtgc agtcggcttc tgacgttcag tgcagccgtc ttctgaaaac
1381 gacatgtcgc acaagtccta agttacgcga caggctgccg ccctgccctt ttcctggcgt
1441 tttcttgtcg cgtgttttag tcgcataaag tagaatactt gcgactagaa ccggagacat
1501 tacgccatga acaagagcgc cgccgctggc ctgctgggct atgcccgcgt cagcaccgac
1561 gaccaggact tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac caagctgttt
1621 tccgagaaga tcaccggcac caggcgcgac cgcccggagc tggccaggat gcttgaccac
1681 ctacgccctg gcgacgttgt gacagtgacc aggctagacc gcctggcccg cagcacccgc
1741 gacctactgg acattgccga gcgcatccag gaggccggcg cgggcctgcg tagcctggca
1801 gagccgtggg ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc
1861 attgccgagt tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc
1921 aaggcccgag gcgtgaagtt tggcccccgc cctaccctca ccccggcaca gatcgcgcac
1981 gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag aggcggctgc actgcttggc
2041 gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag
2101 gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc cctggcggcc
2161 gccgagaatg aacgccaaga ggaacaagca tgaaaccgca ccaggacggc caggacgaac
2221 cgtttttcat taccgaagag atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc
2281 cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca
2341 agctggcggc ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa
2401 ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat
2461 gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa
2521 aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg
2581 ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt
2641 gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt
2701 gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt
2761 ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta
2821 cgacatatgg gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga
2881 tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg
2941 tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca
3001 gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga
3061 gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg
3121 agttaatgag gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg
3181 agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag
3241 cgggtcaact ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc
3301 caaggcaaga ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg
3361 agcaaatgaa taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca
3421 agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc
3481 aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc actggaaccc ccaagcccga
3541 ggaatcggcg tgacggtcgc aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg
3601 atgacctggt ggagaagttg aaggccgcgc aggccgccca gcggcaacgc atcgaggcag
3661 aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc
3721 aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc caagggcgac gagcaaccag
3781 attttttcgt tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc atcatggacg
3841 tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc
3901 ttccagacgg gcacgtagag gtttccgcag ggccggccgg catggccagt gtgtgggatt
3961 acgacctggt actgatggcg gtttcccatc taaccgaatc catgaaccga taccgggaag
4021 ggaagggaga caagcccggc cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct
4081 gccggcgagc cgatggcgga aagcagaaag acgacctggt agaaacctgc attcggttaa
4141 acaccacgca cgttgccatg cagcgtacga agaaggccaa gaacggccgc ctggtgacgg
4201 tatccgaggg tgaagccttg attagccgct acaagatcgt aaagagcgaa accgggcggc
4261 cggagtacat cgagatcgag ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga
4321 acccggacgt gctgacggtt caccccgatt actttttgat cgatcccggc atcggccgtt
4381 ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga
4441 cgatctacga acgcagtggc agcgccggag agttcaagaa gttctgtttc accgtgcgca
4501 agctgatcgg gtcaaatgac ctgccggagt acgatttgaa ggaggaggcg gggcaggctg
4561 gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg cgaagcatcc gccggttcct
4621 aatgtacgga gcagatgcta gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggtc
4681 tctttcctgt ggatagcacg tacattggga acccaaagcc gtacattggg aaccggaacc
4741 cgtacattgg gaacccaaag ccgtacattg ggaaccggtc acacatgtaa gtgactgata
4801 taaaagagaa aaaaggcgat ttttccgcct aaaactcttt aaaacttatt aaaactctta
4861 aaacccgcct ggcctgtgca taactgtctg gccagcgcac agccgaagag ctgcaaaaag
4921 cgcctaccct tcggtcgctg cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg
4981 ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa tctaccaggg cgcggacaag
5041 ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc
5101 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
5161 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
5221 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
5281 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
5341 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
5401 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
5461 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
5521 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
5581 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
5641 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
5701 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
5761 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
5821 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
5881 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
5941 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
6001 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
6061 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
6121 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
6181 ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc taggtactaa aacaattcat
6241 ccagtaaaat ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa
6301 atagctcgac atactgttct tccccgatat cctccctgat cgaccggacg cagaaggcaa
6361 tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc
6421 catctttcac aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt
6481 cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtgtctt
6541 cttcccagtt ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg
6601 ctaagcggct gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga
6661 gcctgatgca ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact
6721 cttccgagca aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc
6781 gttcaaagtg caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct
6841 tttcccgttc cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata
6901 ggttttcatt ttctcccacc agcttatata ccttagcagg agacattcct tccgtatctt
6961 ttacgcagcg gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca
7021 tttattattt ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa
7081 caagacgaac tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc
7141 tttttcaaag ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa
7201 accgcggtga tcacaggcag caacgctctg tcatcgttac aatcaacatg ctaccctccg
7261 cgagatcatc cgtgtttcaa acccggcagc ttagttgccg ttcttccgaa tagcatcggt
7321 aacatgagca aagtctgccg ccttacaacg gctctcccgc tgacgccgtc ccggactgat
7381 gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg
7441 gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca
7501 ttgcggacgt ttttaatgta ctgaattaac gccgaattaa ttcgggggat ctggatttta
7561 gtactggatt ttggttttag gaattagaaa ttttattgat agaagtattt tacaaataca
7621 aatacatact aagggtttct tatatgctca acacatgagc gaaaccctat aggaacccta
7681 attcccttat ctgggaacta ctcacacatt attatggaga aactcgagct tgtcgatcga
7741 cagatccggt cggcatctac tctatttctt tgccctcgga cgagtgctgg ggcgtcggtt
7801 tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc ttctgcgggc
7861 gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc atcgaccctg
7921 cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt ggtcaagacc
7981 aatgcggagc atatacgccc ggagtcgtgg cgatcctgca agctccggat gcctccgctc
8041 gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga agatgttggc
8101 gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg ctgttatgcg
8161 gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt gccggacttc
8221 ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg acgcactgac
8281 ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg cgcatatgaa
8341 atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga acccgctcgt
8401 ctggctaaga tcggccgcag cgatcgcatc catagcctcc gcgaccggtt gtagaacagc
8461 gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc gggagatgca
8521 ataggtcagg ctctcgctaa actccccaat gtcaagcact tccggaatcg ggagcgcggc
8581 cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc agctatttac
8641 ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt cttcgccctc
8701 cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact tctcgacaga
8761 cgtcgcggtg agttcaggct ttttcatatc tcattgcccc ccgggatctg cgaaagctcg
8821 agagagatag atttgtagag agagactggt gatttcagcg tgtcctctcc aaatgaaatg
8881 aacttcctta tatagaggaa ggtcttgcga aggatagtgg gattgtgcgt catcccttac
8941 gtcagtggag atatcacatc aatccacttg ctttgaagac gtggttggaa cgtcttcttt
9001 ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg cagaggcatc
9061 ttgaacgata gcctttcctt tatcgcaatg atggcatttg taggtgccac cttccttttc
9121 tactgtcctt ttgatgaagt gacagatagc tgggcaatgg aatccgagga ggtttcccga
9181 tattaccctt tgttgaaaag tctcaatagc cctttggtct tctgagactg tatctttgat
9241 attcttggag tagacgagag tgtcgtgctc caccatgtta tcacatcaat ccacttgctt
9301 tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg ggggtccatc
9361 tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat cgcaatgatg
9421 gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac agatagctgg
9481 gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct caatagccct
9541 ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt cgtgctccac
9601 catgttggca agctgctcta gccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
9661 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
9721 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
9781 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
9841 gattacg
//