LOCUS Exported 6968 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION GoldenBraid level 1 Alpha plant expression vector. . ACCESSION . VERSION . KEYWORDS pDGB3_alpha2R SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6968) 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 6968) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6968 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 87..108 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 123..153 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 161..177 /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)." primer_bind 166..188 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 185..201 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 185..201 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" misc_feature 214..270 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(271..288) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(271..287) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(280..302) /label=M13/pUC Forward /note="In lacZ gene" misc_feature 634..658 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 1959..2588 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 3022..4089 /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 4155..4349 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(4585..4607) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 4693..4833 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4848..4865) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5019..5607) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5099..5118) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5695..6510) /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 6418..6437 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" misc_feature 6943..6967 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" ORIGIN 1 taacgatatc gtctcagtca agcgagagac cccagctggc acgacaggtt tcccgactgg 61 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag 121 gctttacact atatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt 181 cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct 241 agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg 301 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg 361 gcgtaatagc gaagaggcct gcgccgatcg cccttcccaa cagttgcgca gcctgaatgg 421 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat 481 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc 541 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 601 agctgtgggt ctctctccgc ttgagacgat atctgacagg atatattggc gggtaaacct 661 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc 721 cgttcgtcca tttgtatgtg catgccaacc acagggttcc cctcgggatc aaagtacttt 781 gatccaaccc ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcatc 841 tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc 901 ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg 961 gagacattac gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag 1021 caccgacgac caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa 1081 gctgttttcc gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct 1141 tgaccaccta cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag 1201 cacccgcgac ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag 1261 cctggcagag ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt 1321 cgccggcatt gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga 1381 ggccgccaag gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat 1441 cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact 1501 gcttggcgtg catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc 1561 caccgaggcc aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct 1621 ggcggccgcc gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag 1681 gacgaaccgt ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg 1741 ttcgagccgc ccgcgcacct ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct 1801 gatgccaagc tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt 1861 ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg 1921 atgcgatgag taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta 1981 accagaaagg cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac 2041 tcgccggggc cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg 2101 cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc 2161 gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg 2221 cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa 2281 gcccttacga catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg 2341 tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca 2401 tcggcggtga ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta 2461 tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag 2521 aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac 2581 tcatttgagt taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg 2641 ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc 2701 catgaagcgg gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc 2761 ggtacgccaa ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga 2821 gtaaatgagc aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg 2881 aaaatcaaga acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg 2941 gttggccagg cgtaagcggc tgggttgtct gccggccctg caatggcact ggaaccccca 3001 agcccgagga atcggcgtga cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg 3061 ctgggtgatg acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc 3121 gaggcagaag cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa 3181 tcccggcaac cgccggcagc cggtgcgccg tcgattagga agccgcccaa gggcgacgag 3241 caaccagatt ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc 3301 atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc 3361 tacgagcttc cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg 3421 tgggattacg acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac 3481 cgggaaggga agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc 3541 aagttctgcc ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt 3601 cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg 3661 gtgacggtat ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc 3721 gggcggccgg agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa 3781 ggcaagaacc cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc 3841 ggccgttttc tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg 3901 ttcaagacga tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc 3961 gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg 4021 caggctggcc cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc 4081 ggttcctaat gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga 4141 aaaggactct ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac 4201 cggaacccgt acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg 4261 actgatataa aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa 4321 actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg 4381 caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct 4441 atcgcggccg ctggccgctc aaaaatggct ggcctacggc caggcaatct accagggcgc 4501 ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac atcaaggcac cctgcctcgc 4561 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggtga cggtcacagc 4621 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 4681 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt 4741 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg 4801 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac 4861 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 4921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 4981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 5041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 5221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 5281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 5341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 5401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 5461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 5521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 5581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 5641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcattctag gtgattagaa 5701 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata 5761 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat 5821 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa 5881 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc 5941 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt 6001 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg 6061 agcgagtcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa 6121 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc 6181 taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg 6241 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct 6301 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc 6361 tggcgcatcg ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc 6421 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga 6481 gcaagacgtt tcccgttgaa tatggctcat aacagaactt attatttcct tcctcttttc 6541 tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt 6601 tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt 6661 tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa 6721 cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc 6781 cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct 6841 tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga 6901 ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg 6961 tgtaaaca //