LOCUS Exported 6967 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION GoldenBraid level 1 Alpha plant expression vector. . ACCESSION . VERSION . KEYWORDS pDGB3_alpha2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6967) AUTHORS Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D TITLE GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. JOURNAL Plant Physiol. 2013 Jul;162(3):1618-31. PUBMED 23669743 REFERENCE 2 (bases 1 to 6967) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6967 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 338..360 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 352..369 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 353..369 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 370..426 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(439..455) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(439..455) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(452..474) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 463..479 /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)." promoter complement(487..517) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 532..553 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." misc_feature 633..657 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 1958..2587 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 3021..4088 /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 4154..4348 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(4584..4606) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 4692..4832 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4847..4864) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5018..5606) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5098..5117) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5694..6509) /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" primer_bind 6417..6436 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" misc_feature 6942..6966 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" ORIGIN 1 taacaagctt cgtctcagtc aggagagaga cccacagctt gtctgtaagc ggatgccggg 61 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac 121 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac 181 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt 241 tgggaagggc gatcggcgca ggcctcttcg ctattacgcc agctggcgaa agggggatgt 301 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 361 acggccagtg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc 421 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 481 tccacacaac atacgagccg gaagcatata gtgtaaagcc tggggtgcct aatgagtgag 541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 601 ccagctgggg tctctcgctt gagacgaagc tttgacagga tatattggcg ggtaaaccta 661 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc 721 gttcgtccat ttgtatgtgc atgccaacca cagggttccc ctcgggatca aagtactttg 781 atccaacccc tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcatct 841 gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc 901 tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg 961 agacattacg ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc 1021 accgacgacc aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag 1081 ctgttttccg agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt 1141 gaccacctac gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc 1201 acccgcgacc tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc 1261 ctggcagagc cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc 1321 gccggcattg ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag 1381 gccgccaagg cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc 1441 gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg 1501 cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc 1561 accgaggcca ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg 1621 gcggccgccg agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg 1681 acgaaccgtt tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt 1741 tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg 1801 atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc 1861 taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga 1921 tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa 1981 ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact 2041 cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc 2101 ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg 2161 cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc 2221 ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag 2281 cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt 2341 cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat 2401 cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat 2461 cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga 2521 acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact 2581 catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc 2641 cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc 2701 atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg 2761 gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag 2821 taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga 2881 aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg 2941 ttggccaggc gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa 3001 gcccgaggaa tcggcgtgac ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc 3061 tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg 3121 aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat 3181 cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc 3241 aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca 3301 tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct 3361 acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt 3421 gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc 3481 gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca 3541 agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc 3601 ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg 3661 tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg 3721 ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag 3781 gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg 3841 gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt 3901 tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg 3961 tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc 4021 aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg 4081 gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa 4141 aaggactctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc 4201 ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga 4261 ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa 4321 ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc 4381 aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta 4441 tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg 4501 gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg 4561 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggtgac ggtcacagct 4621 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc 4681 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta 4741 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc 4801 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact 4861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 4921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 4981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 5041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 5101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 5161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 5221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 5281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 5341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 5401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga 5461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 5521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 5581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcattctagg tgattagaaa 5701 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat 5761 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg 5821 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat 5881 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc 5941 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta 6001 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga 6061 gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac 6121 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct 6181 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga 6241 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg 6301 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct 6361 ggcgcatcgg gcttcccata caatcggtag attgtcgcac ctgattgccc gacattatcg 6421 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag 6481 caagacgttt cccgttgaat atggctcata acagaactta ttatttcctt cctcttttct 6541 acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt 6601 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt 6661 ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac 6721 gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc 6781 ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt 6841 acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat 6901 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt 6961 gtaaaca //