LOCUS Exported 8958 bp ds-DNA circular SYN 13-SEP-2016
DEFINITION synthetic circular DNA
KEYWORDS Untitled 7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8958)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8958
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(60..76)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 279..303
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS 1603..2232
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 2666..3733
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/note="pVS1 RepA"
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 3799..3993
/note="pVS1 oriV"
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 4337..4477
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4663..5251)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5338..6132)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 6557..6581
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 6659..6833
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(6873..7898)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(7966..8643)
/note="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 8834..8855
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8870..8900
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 8908..8924
/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)."
primer_bind 8932..8948
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 8944..219
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
misc_feature 8958..56
/note="MCS"
/note="pUC18/19 multiple cloning site"
ORIGIN
1 aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggca
61 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
121 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
181 ccttcccaac agttgcgcag cctgaatggc gaatgctaga gcagcttgag cttggatcag
241 attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata ttggcgggta
301 aacctaagag aaaagagcgt ttattagaat aacggatatt taaaagggcg tgaaaaggtt
361 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta
421 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt
481 cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct
541 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga
601 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg
661 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca
721 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga
781 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc
841 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc
901 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg
961 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc
1021 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac
1081 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg
1141 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga
1201 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg
1261 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg
1321 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta
1381 cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc tggccggttt
1441 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
1501 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
1561 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
1621 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
1681 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
1741 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
1801 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
1861 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
1921 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt
1981 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg
2041 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc
2101 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa
2161 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca
2221 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg
2281 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc
2341 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt
2401 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac
2461 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc
2521 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg
2581 ggcggttggc caggcgtaag cggctgggtt gtctgccggc cctgcaatgg cactggaacc
2641 cccaagcccg aggaatcggc gtgacggtcg caaaccatcc ggcccggtac aaatcggcgc
2701 ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg
2761 catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa
2821 agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga
2881 cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag
2941 catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat
3001 ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag
3061 tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg
3121 ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt
3181 actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg
3241 cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg
3301 cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga
3361 aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac
3421 agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg
3481 catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg
3541 gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt
3601 caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc
3661 ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc
3721 cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg
3781 tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg
3841 gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta
3901 agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat
3961 taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga
4021 gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg
4081 gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg
4141 gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc
4201 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
4261 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
4321 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
4381 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
4441 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
4501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
4561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
4621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
4681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
4741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
4801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
4861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
4921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
4981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
5041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
5101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
5161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
5221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
5281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatt ctaggtacta
5341 aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct tgatccccag
5401 taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga tcgaccggac
5461 gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat caataaagcc
5521 acttactttg ccatctttca caaagatgtt gctgtctccc aggtcgccgt gggaaaagac
5581 aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa
5641 tggagtgtct tcttcccagt tttcgcaatc cacatcggcc agatcgttat tcagtaagta
5701 atccaattcg gctaagcggc tgtctaagct attcgtatag ggacaatccg atatgtcgat
5761 ggagtgaaag agcctgatgc actccgcata cagctcgata atcttttcag ggctttgttc
5821 atcttcatac tcttccgagc aaaggacgcc atcggcctca ctcatgagca gattgctcca
5881 gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt ccagccatag
5941 catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc tgtccgtcat
6001 ttttaaatat aggttttcat tttctcccac cagcttatat accttagcag gagacattcc
6061 ttccgtatct tttacgcagc ggtatttttc gatcagtttt ttcaattccg gtgatattct
6121 cattttagcc atttattatt tccttcctct tttctacagt atttaaagat accccaagaa
6181 gctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa ccttaaatac
6241 cagaaaacag ctttttcaaa gttgttttca aagttggcgt ataacatagt atcgacggag
6301 ccgattttga aaccgcggtg atcacaggca gcaacgctct gtcatcgtta caatcaacat
6361 gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga
6421 atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt
6481 cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga
6541 gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac
6601 ttaataacac attgcggacg tttttaatgt actgaattaa cgccgaatta attcggggga
6661 tctggatttt agtactggat tttggtttta ggaattagaa attttattga tagaagtatt
6721 ttacaaatac aaatacatac taagggtttc ttatatgctc aacacatgag cgaaacccta
6781 taggaaccct aattccctta tctgggaact actcacacat tattatggag aaactcgagc
6841 ttgtcgatcg acagatccgg tcggcatcta ctctatttct ttgccctcgg acgagtgctg
6901 gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg
6961 cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg
7021 catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt
7081 tggtcaagac caatgcggag catatacgcc cggagtcgtg gcgatcctgc aagctccgga
7141 tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag
7201 aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc
7261 gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg
7321 tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg
7381 gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc
7441 gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg
7501 aacccgctcg tctggctaag atcggccgca gcgatcgcat ccatagcctc cgcgaccggt
7561 tgtagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg
7621 cgggagatgc aataggtcag gctctcgcta aactccccaa tgtcaagcac ttccggaatc
7681 gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg
7741 cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat
7801 tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac
7861 ttctcgacag acgtcgcggt gagttcaggc tttttcatat ctcattgccc cccgggatct
7921 gcgaaagctc gagagagata gatttgtaga gagagactgg tgatttcagc gtgtcctctc
7981 caaatgaaat gaacttcctt atatagagga aggtcttgcg aaggatagtg ggattgtgcg
8041 tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga cgtggttgga
8101 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
8161 gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt gtaggtgcca
8221 ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg gaatccgagg
8281 aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact
8341 gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt atcacatcaa
8401 tccacttgct ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt
8461 gggggtccat ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta
8521 tcgcaatgat ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga
8581 cagatagctg ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc
8641 tcaatagccc tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg
8701 tcgtgctcca ccatgttggc aagctgctct agccaatacg caaaccgcct ctccccgcgc
8761 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
8821 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
8881 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
8941 gctatgacca tgattacg
//
Caution:
Product is for research use only!
Search name
pCAMBIA1300,Plasmid pCAMBIA1300,pCAMBIA1300 vector
English