LOCUS Exported 7448 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION GoldenBraid level alpha luciferase reporter. ACCESSION . VERSION . KEYWORDS pEPYC-alpha-2rKN0009 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7448) 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 7448) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7448 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(1226..1855) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIGTHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" misc_feature 3156..3180 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 3403..3912 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW RLCERIL" CDS 3976..3999 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 4015..4267 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_feature 4295..4319 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 4752..5567 /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 complement(4825..4844) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 5655..6243 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6144..6163 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6397..6414 /label=L4440 /note="L4440 vector, forward primer" misc_feature 6429..6569 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(6495..6514) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6655..6677 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 6913..7107 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(7173..792) /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" ORIGIN 1 ggggtgaacc gtcagcacgt ccgggttctt gccttctgtg atctcgcggt acatccaatc 61 agctagctcg atctcgatgt actccggccg cccggtttcg ctctttacga tcttgtagcg 121 gctaatcaag gcttcaccct cggataccgt caccaggcgg ccgttcttgg ccttcttcgt 181 acgctgcatg gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt 241 ctgctttccg ccatcggctc gccggcagaa cttgagtacg tccgcaacgt gtggacggaa 301 cacgcggccg ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt cggttagatg 361 ggaaaccgcc atcagtacca ggtcgtaatc ccacacactg gccatgccgg ccggccctgc 421 ggaaacctct acgtgcccgt ctggaagctc gtagcggatc acctcgccag ctcgtcggtc 481 acgcttcgac agacggaaaa cggccacgtc catgatgctg cgactatcgc gggtgcccac 541 gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat 601 cgacggcgca ccggctgccg gcggttgccg ggattctttg cggattcgat cagcggccgc 661 ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc 721 ggccttcaac ttctccacca ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga 781 tggtttgcga ccgtcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc 841 ggcagacaac ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt 901 ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa 961 ttcatctact catttattca tttgctcatt tactctggta gctgcgcgat ctattcagat 1021 agcagctcgg taatggtctt gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc 1081 tccgccggca actgaaagtt gacccgcttc atggctggcg tgtctgccag gctggccaac 1141 gttgcagcct tgctgctgcg tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg 1201 ctcattttct ctttacctca ttaactcaaa tgagttttga tttaatttca gcggccagcg 1261 cctggacctc gcgggcagcg tcgccctcgg gttctgattc aagaacggtt gtgccggcgg 1321 cggcagtgcc tgggtagctc acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt 1381 acccggccag cgcctcggca acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca 1441 cgacaaaggc cgcttgtagc cttccatccg tgacctcaat gcgctgctta accagctcca 1501 ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg caccggaatc agcacgaagt 1561 cggctgcctt gatcgcggac acagccaagt ccgccgcctg gggcgctccg tcgatcacta 1621 cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga 1681 caacggttag cggttgatct tcccgcacgg ccgcccaatc gcgggcactg ccctggggat 1741 cggaatcgac taacagaaca tcggccccgg cgagttgcag ggcgcgggct agatgggttc 1801 cgatggtcgt cttgcctgac ccgcctttct ggttaagtac agcgataacc ttcatgcgtt 1861 ccccttgcgt atttgtttat ttactcatcg gatcatatac gcagcgaccg catgacgcaa 1921 gctgttttac tcaaatacac atcacctttt tagacggcgg cgctcggttt cttcagcggc 1981 caagctggcc ggccaggccg ccagcttggc atcagacaaa ccggccagga tttcatgcag 2041 ccgcacggtt gagaggtgcg cgggcggctc gaacacgtac ccggccgcga tcatctccgc 2101 ctcgatctct tcggtaatga aaaacggttc gtcctggccg tcctggtgcg gtttcatgct 2161 tgttcctctt ggcgttcatt ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc 2221 tcacggaagg caccgcgccg cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt 2281 gcgcggtaca gggtcgagcg atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg 2341 ccttcctggt cgatcagctc gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg 2401 gggccaaact tcacgcctcg ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg 2461 attagggaac gctcgaactc ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc 2521 ggcgtggtgg tgtcggccca cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg 2581 atgcgctcgg caatgtccag taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc 2641 actgtcacaa cgtcgccagg gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg 2701 cgcctggtgc cggtgatctt ctcggaaaac agcttggtgc agccggccgc gtgcagttcg 2761 gcccgttggt tggtcaagtc ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca 2821 gcggcggcgc tcttgttcat ggcgtaatgt ctccggttct agtcgcaagt attctacttt 2881 atgcgactaa aacacgcgac aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc 2941 gtaacttagg acttgtgcga catgtcgttt tcagatgacg gctgcactga acgtcagaag 3001 ccgactgcac tatagcagcg gaggggttgg atcaaagtac tttgatcccg aggggaaccc 3061 tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat 3121 atccgattat tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca 3181 gatatcgtct caagcggagg ttatcacact tggtaattta gaagaagaaa cccataggtt 3241 ctgtgggttt tttgggtaga agaagaaacc cataggttct gtgggttttt tgggtagaag 3301 aagaaaccca taggttctgt gggttttttg ggtagaagaa gaaacccata gctatataat 3361 gagaaacagg ggcaaatccc cgaaaacaaa taacgcgaag caatggtatt cacactcgaa 3421 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag 3481 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt 3541 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt 3601 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat 3661 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg 3721 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag 3781 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac 3841 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc 3901 gaacgcattc tggcttcgga ttataaggac catgacggag actataagga ccatgacctc 3961 gacgctgcag cagcggatta taaggacgat gacgataagt gagcttgtca agcagatcgt 4021 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt 4081 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg 4141 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata 4201 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta 4261 ctagatcgac gcttgactga gacgatatcg ttatgtttac accacaatat atcctgccac 4321 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca 4381 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca 4441 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat 4501 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaccg 4561 cggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga 4621 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt 4681 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa 4741 taagttctgt tatgagccat attcaacggg aaacgtcttg ctcaaggccg cgattaaatt 4801 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag 4861 gtgcgacaat ctaccgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg 4921 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg 4981 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac 5041 tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat cctgattcag 5101 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt 5161 gtaattgtcc ttttaacagc gatcgcgtat ttcgactcgc tcaggcgcaa tcacgaatga 5221 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac 5281 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg 5341 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg 5401 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg 5461 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg 5521 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca cctagaatgc 5581 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 5641 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 5701 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 5761 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 5821 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 5881 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 5941 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 6001 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 6061 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 6121 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 6181 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 6241 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 6301 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 6361 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 6421 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 6481 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc 6541 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc 6601 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtcaccggga 6661 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ggtgccttga 6721 tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg tagattgcct 6781 ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg cgaagcggcg 6841 gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc ggctgtgcgc 6901 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag 6961 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg 7021 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt 7081 gggttcccaa tgtacgtgct atccacagga aagagtcctt ttcgaccttt ttcccctgct 7141 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg 7201 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa 7261 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg 7321 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc 7381 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa 7441 aagtaatc //