LOCUS Exported 7286 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION GoldenBraid level alpha luciferase reporter. ACCESSION . VERSION . KEYWORDS pEPKK-alpha-2KN0091 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7286) AUTHORS Cai YM, Kallam K, Tidd H, Gendarini G, Salzman A, Patron NJ TITLE Rational design of minimal synthetic promoters for plants. JOURNAL Nucleic Acids Res. 2020 Aug 28. pii: 5897334. doi: 10.1093/nar/gkaa682. PUBMED 32856047 REFERENCE 2 (bases 1 to 7286) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7286 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 183..207 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 319..828 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW RLCERIL" CDS 892..915 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 938..1138 /label=gene 7 terminator /note="transcription terminator for octopine-type Ti plasmid gene 7" misc_feature 1160..1184 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 2485..3114 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 3548..4615 /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 4681..4875 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(5111..5133) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 5219..5359 /label=bom /note="basis of mobility region from pBR322" primer_bind 5274..5293 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5374..5391) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5545..6133) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5625..5644) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6221..7036) /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 6944..6963 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" ORIGIN 1 gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac 61 atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg 121 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct 181 ggtggcagga tatattgtgg tgtaaacata acaagcttcg tctcaggagg ttctgtgggt 241 tttttgggta gaagaagaaa cccatagcta tataatgaga aacaggggca aatccccgaa 301 aacaaataac gcgaagcaat ggtattcaca ctcgaagatt tcgttgggga ctggcgacag 361 acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgtttcag 421 aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg 481 aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag 541 atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg 601 cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg 661 ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg 721 aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga 781 gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc ttcggattat 841 aaggaccatg acggagacta taaggaccat gacctcgacg ctgcagcagc ggattataag 901 gacgatgacg ataagtgagc ttgatccccc gtcgacagct agctatatca tcaatttatg 961 tattacacat aatatcgcac tcagtctttc atctacggca atgtaccagc tgatataatc 1021 agttattgaa atatttctga atttaaactt gcatcaataa atttatgttt ttgcttggac 1081 tataatacct gacttgttat tttatcaata aatatttaaa ctatatttct ttcaagatcg 1141 ctgtcatgag acgaagcttt gacaggatat attggcgggt aaacctaaga gaaaagagcg 1201 tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg 1261 tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc caacccctcc 1321 gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcatctgaa aacgacatgt 1381 cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg 1441 tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca 1501 tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg 1561 acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga 1621 agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc 1681 ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac 1741 tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt 1801 gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg 1861 agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc 1921 gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg 1981 agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc 2041 gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc 2101 ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga 2161 atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt 2221 cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc 2281 gcacctctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc 2341 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg 2401 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatcc gatgagtaaa 2461 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg 2521 tcaggcaaga cgaccatcgg aacccatcta gcccgcgccc tgcaactcgc cggggccgat 2581 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa 2641 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc 2701 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg 2761 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata 2821 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg 2881 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt 2941 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg 3001 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac 3061 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat 3121 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac 3181 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca 3241 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca 3301 agaccattac cgagctgcta tctgaataga tcgcgcagct accagagtaa atgagcaaat 3361 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa 3421 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta 3481 agcggctggg ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg 3541 gcgtgacggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg gtgatgacct 3601 ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg cagaagcacg 3661 ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc ggcaaccgcc 3721 ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac cagatttttt 3781 cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg acgtggccgt 3841 tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg agcttccaga 3901 cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg attacgacct 3961 ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg aagggaaggg 4021 agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt tctgccggcg 4081 agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt taaacaccac 4141 gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga cggtatccga 4201 gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc ggccggagta 4261 catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca agaacccgga 4321 cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta 4381 ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta 4441 cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat 4501 cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat 4561 cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac 4621 ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gactctttcc 4681 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat 4741 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga 4801 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg 4861 cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac 4921 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg 4981 ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc 5041 gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg 5101 acggtgaaaa cctctgacac atgcagctcc cggtgacggt cacagcttgt ctgtaagcgg 5161 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg 5221 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc 5281 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 5341 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat 6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 6061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 6181 aaactcacgt taagggattt tggtcatgca ttctaggtga ttagaaaaac tcatcgagca 6241 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc 6301 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt 6361 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa 6421 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca 6481 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa 6541 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agtcgaaata 6601 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca 6661 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg 6721 ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat 6781 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg 6841 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct 6901 tcccatacaa tcggtagatt gtcgcacctg attgcccgac attatcgcga gcccatttat 6961 acccatataa atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc 7021 gttgaatatg gctcataaca gaacttatta tttccttcct cttttctaca gtatttaaag 7081 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa 7141 aaccttaaat accagaaaac agctttttca aagttgtttt caaagttggc gtataacata 7201 gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg cagcaacgct ctgtcatcgt 7261 tacaatcaac atgctaccct ccgcga //