LOCUS Exported 7332 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo Level 1 luciferase reporter. ACCESSION . VERSION . KEYWORDS pEPYC1CB0127 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7332) 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 7332) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7332 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 42..66 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS complement(222..1370) /codon_start=1 /product="trans-acting replication protein that binds to and activates oriV" /label=trfA /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" rep_origin complement(1706..2294) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1786..1805) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2465..3325) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3088..3107 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3326..3430) /gene="bla" /label=AmpR promoter rep_origin 3517..4148 /label=oriV /note="incP origin of replication" misc_feature 4240..4264 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" primer_bind 4738..4758 /label=35S promoter /note="CaMV 35S promoter, forward primer" promoter 4747..4793 /label=minimal CaMV 35S promoter /note="minimal 35S promoter from cauliflower mosaic virus" misc_feature 4814..4869 /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" CDS 4871..6520 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(4937..4955) /label=LucNrev /note="Firefly luciferase, reverse primer" CDS 6584..6607 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 6618..7325 /label=OCS terminator /note="octopine synthase terminator" ORIGIN 1 gcaattgtct tctgcacgaa gtggtttaaa ctatcagtgt ttgacaggat atattggcgg 61 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa 121 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc agatcaggcg 181 ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc 241 aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct 301 tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg 361 aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt 421 cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca 481 gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca 541 cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg 601 gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa 661 taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc 721 ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca 781 tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa 841 atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca 901 gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac 961 aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg 1021 gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata 1081 gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcaagacga 1141 cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg 1201 tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg gaaggtttcg 1261 cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc 1321 gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct 1381 tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt tgacccgcct 1441 ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc gtagaccgtc 1501 tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac cagcgacccc 1561 ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat gcggaagaag 1621 tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggatct gccaggaacc 1681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 1741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 1801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 1861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 1921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 1981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 2041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 2101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 2161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 2221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 2281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 2341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 2581 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa 2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 2761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 3061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 3361 aaggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 3421 gggttccgcg cacgaattgg ccagcgctgc catttttggg gtgaggccgt tcgcggccga 3481 ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag 3541 ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa 3601 caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa 3661 acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca 3721 ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc 3781 atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg 3841 tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg 3901 gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg 3961 gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt 4021 ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc acacggcttc gacggcgttt 4081 ctggcgcgtt tgcagggcca tagacggccg ccagcccagc ggcgagggca accagcccgg 4141 tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg cctgtatcga gtggtgattt 4201 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt 4261 aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg 4321 tggatgcagt gggccccact ctgtgaagac aatgccgaat tcggatccgg aggtcaacat 4381 ggtggagcac gacactctgg tctactccaa aaatgtcaaa gatacagtct cagaagatca 4441 aagggctatt gagacttttc aacaaaggat aatttcggga aacctcctcg gattccattg 4501 cccagctatc tgtcacttca tcgaaaggac agtagaaaag gaaggtggct cctacaaatg 4561 ccatcattgc gataaaggaa aggctatcat tcaagactga cgtaagggat gacgcacaga 4621 tggaccccca cccacgagga gcatcgtgga aaaagaagag gttcctctct ctgccgacag 4681 tggtcccaaa aaccacgtct acaatgatgt gacatctcca agcaagtgga tatcccacta 4741 tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga ggacacgcta 4801 ctgtattttt acaacaatta ccaacaacaa caaacaacaa acaacattac aattactatt 4861 tacaattaca atggaggacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct 4921 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc 4981 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt 5041 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag 5101 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt 5161 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag 5221 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt 5281 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga 5341 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa 5401 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa 5461 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt 5521 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat 5581 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat 5641 atgtggattt cgagtcgtct taatgtatag atttgaagaa gaactgtttc tgaggagcct 5701 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa 5761 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc 5821 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag 5881 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga 5941 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga 6001 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat 6061 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg 6121 gctacattct ggagacatag cttactggga cgaggacgaa cacttcttca tcgttgaccg 6181 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat 6241 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtctgcccg acgatgacgc 6301 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga 6361 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt 6421 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga 6481 gatcctcata aaggccaaga agggcggaaa gatcgccgtg ggttcggatt ataaggacca 6541 tgacggagac tataaggacc atgacctcga cgctgcagca gcggattata aggacgatga 6601 cgataagtga gcttgtcctg ctttaatgag atatgcgaga agcctatgat cgcatgatat 6661 ttgctttcaa ttctgttgtg cacgttgtaa aaaacctgag catgtgtagc tcagatcctt 6721 accgccggtt tcggttcatt ctaatgaata tatcacccgt tactatcgta tttttatgaa 6781 taatattctc cgttcaattt actgattgta ccctactact tatatgtaca atattaaaat 6841 gaaaacaata tattgtgctg aataggttta tagcgacatc tatgatagag cgccacaata 6901 acaaacaatt gcgttttatt attacaaatc caattttaaa aaaagcggca gaaccggtca 6961 aacctaaaag actgattaca taaatcttat tcaaatttca aaagtgcccc aggggctagt 7021 atctacgaca caccgagcgg cgaactaata acgctcactg aagggaactc cggttccccg 7081 ccggcgcgca tgggtgagat tccttgaagt tgagtattgg ccgtccgctc taccgaaagt 7141 tacgggcacc attcaacccg gtccagcacg gcggccgggt aaccgacttg ctgccccgag 7201 aattatgcag catttttttg gtgtatgtgg gccccaaatg aagtgcaggt caaaccttga 7261 cagtgacgac aaatcgttgg gcgggtccag ggcgaatttt gcgacaacat gtcgaggctc 7321 agcaggaccg ct //