LOCUS Exported 8428 bp ds-DNA circular SYN 14-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCAMBIA3300
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8428)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Wednesday, September 14, 2016 from SnapGene Viewer 3.2.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8428
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(38..715)
/note="CaMV 35S promoter (enhanced)"
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 906..927
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 942..972
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 980..996
/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 1004..1020
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 1016..1249
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
misc_feature 1030..1086
/note="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(1090..1106)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1309..1333
/note="RB T-DNA repeat"
/note="right border repeat from nopaline C58 T-DNA"
CDS 2633..3262
/codon_start=1
/product="stability protein from Pseudomonas plasmid pVS1"
/note="pVS1 StaA"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 3696..4763
/codon_start=1
/product="replication protein from Pseudomonas plasmid
pVS1"
/note="pVS1 RepA"
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
misc_feature 5367..5507
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5693..6281)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6368..7162)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 7587..7611
/note="LB T-DNA repeat"
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 7689..7863
/note="CaMV poly(A) signal"
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(7870..8421)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/note="BlpR"
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
ORIGIN
1 tcgagagaga tagatttgta gagagagact ggtgatttca gcgtgtcctc tccaaatgaa
61 atgaacttcc ttatatagag gaaggtcttg cgaaggatag tgggattgtg cgtcatccct
121 tacgtcagtg gagatatcac atcaatccac ttgctttgaa gacgtggttg gaacgtcttc
181 tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagaggc
241 atcttgaacg atagcctttc ctttatcgca atgatggcat ttgtaggtgc caccttcctt
301 ttctactgtc cttttgatga agtgacagat agctgggcaa tggaatccga ggaggtttcc
361 cgatattacc ctttgttgaa aagtctcaat agccctttgg tcttctgaga ctgtatcttt
421 gatattcttg gagtagacga gagtgtcgtg ctccaccatg ttatcacatc aatccacttg
481 ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc
541 atctttggga ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg
601 atggcatttg taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc
661 tgggcaatgg aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc
721 cctttggtct tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc
781 caccatgttg gcaagctgct ctagccaata cgcaaaccgc ctctccccgc gcgttggccg
841 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
901 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
961 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
1021 catgattacg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
1081 aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
1141 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
1201 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgctaga gcagcttgag
1261 cttggatcag attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata
1321 ttggcgggta aacctaagag aaaagagcgt ttattagaat aacggatatt taaaagggcg
1381 tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg
1441 gatcaaagta ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca
1501 gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc
1561 gccctgccct tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact
1621 tgcgactaga accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc
1681 tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg
1741 gccggctgca ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag
1801 ctggccagga tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac
1861 cgcctggccc gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc
1921 gcgggcctgc gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg
1981 gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc
2041 cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc
2101 accccggcac agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa
2161 gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc
2221 gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc
2281 gaggccgacg ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc
2341 accaggacgg ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg
2401 cggccgggta cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc
2461 tggccggttt gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa
2521 ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg
2581 tcgctgcgta tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt
2641 tatcgctgta cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc
2701 ccgcgccctg caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag
2761 tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg
2821 cccgacgatt gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg
2881 agcgccccag gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat
2941 tccggtgcag ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa
3001 gcagcgcatt gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat
3061 caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat
3121 tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac
3181 cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga
3241 aattaaatca aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac
3301 acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg
3361 gcagacacgc cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag
3421 ctgaagatgt acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc
3481 gcgcagctac cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa
3541 aggaggcggc atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg
3601 tggaggaacg ggcggttggc caggcgtaag cggctgggtt gtctgccggc cctgcaatgg
3661 cactggaacc cccaagcccg aggaatcggc gtgacggtcg caaaccatcc ggcccggtac
3721 aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc
3781 agcggcaacg catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc
3841 gaatccgcaa agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc
3901 ccaagggcga cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg
3961 atagtcgcag catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg
4021 gcgaggtgat ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg
4081 gcatggccag tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat
4141 ccatgaaccg ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg
4201 ttgcggacgt actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg
4261 tagaaacctg cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca
4321 agaacggccg cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg
4381 taaagagcga aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc
4441 gcgagatcac agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga
4501 tcgatcccgg catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag
4561 aagccagatg gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga
4621 agttctgttt caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga
4681 aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg
4741 gcgaagcatc cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag
4801 gggaaaaagg tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc
4861 cgtacattgg gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt
4921 cacacatgta agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt
4981 taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca
5041 cagccgaaga gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg
5101 cttcgcgtcg gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca
5161 atctaccagg gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag
5221 gcaccctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
5281 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
5341 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
5401 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
5461 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
5521 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
5581 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
5641 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
5701 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
5761 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
5821 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
5881 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
5941 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
6001 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
6061 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
6121 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
6181 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
6241 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
6301 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatt
6361 ctaggtacta aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct
6421 tgatccccag taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga
6481 tcgaccggac gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat
6541 caataaagcc acttactttg ccatctttca caaagatgtt gctgtctccc aggtcgccgt
6601 gggaaaagac aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg
6661 gatctttaaa tggagtgtct tcttcccagt tttcgcaatc cacatcggcc agatcgttat
6721 tcagtaagta atccaattcg gctaagcggc tgtctaagct attcgtatag ggacaatccg
6781 atatgtcgat ggagtgaaag agcctgatgc actccgcata cagctcgata atcttttcag
6841 ggctttgttc atcttcatac tcttccgagc aaaggacgcc atcggcctca ctcatgagca
6901 gattgctcca gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt
6961 ccagccatag catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc
7021 tgtccgtcat ttttaaatat aggttttcat tttctcccac cagcttatat accttagcag
7081 gagacattcc ttccgtatct tttacgcagc ggtatttttc gatcagtttt ttcaattccg
7141 gtgatattct cattttagcc atttattatt tccttcctct tttctacagt atttaaagat
7201 accccaagaa gctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa
7261 ccttaaatac cagaaaacag ctttttcaaa gttgttttca aagttggcgt ataacatagt
7321 atcgacggag ccgattttga aaccgcggtg atcacaggca gcaacgctct gtcatcgtta
7381 caatcaacat gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc
7441 gttcttccga atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg
7501 ctgacgccgt cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc
7561 cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg
7621 cttagacaac ttaataacac attgcggacg tttttaatgt actgaattaa cgccgaatta
7681 attcggggga tctggatttt agtactggat tttggtttta ggaattagaa attttattga
7741 tagaagtatt ttacaaatac aaatacatac taagggtttc ttatatgctc aacacatgag
7801 cgaaacccta taggaaccct aattccctta tctgggaact actcacacat tattatggag
7861 aaactcgagt caaatctcgg tgacgggcag gaccggacgg ggcggtaccg gcaggctgaa
7921 gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag
7981 catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag
8041 cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt
8101 gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg gagacgtaca cggtcgactc
8161 ggccgtccag tcgtaggcgt tgcgtgcctt ccaggggccc gcgtaggcga tgccggcgac
8221 ctcgccgtcc acctcggcga cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc
8281 cgtccactcc tgcggttcct gcggctcggt acggaagttg accgtgcttg tctcgatgta
8341 gtggttgacg atggtgcaga ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc
8401 cgggcgtcgt tctgggctca tggtagac
//