LOCUS Exported 7041 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo Level 1 luciferase reporter. ACCESSION . VERSION . KEYWORDS pEPYC1CB0435 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7041) 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 7041) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7041 /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" misc_feature 4523..4578 /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" CDS 4580..6229 /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(4646..4664) /label=LucNrev /note="Firefly luciferase, reverse primer" CDS 6293..6316 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 6327..7034 /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 agggaggcgt 4381 gtcgttttag tgaggtcaga agatcaaagg gctatcagaa gatcaaaggg ctatcagaag 4441 atcaaagggc tactatataa ggttttgcta ttcattgaaa gcagtagtga ctgatttgta 4501 tataatgtac tgtattttta caacaattac caacaacaac aaacaacaaa caacattaca 4561 attactattt acaattacaa tggaggacgc caaaaacata aagaaaggcc cggcgccatt 4621 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc 4681 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc 4741 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac 4801 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg 4861 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt 4921 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca 4981 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc 5041 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc 5101 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact 5161 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac 5221 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga 5281 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg 5341 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag aactgtttct 5401 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt 5461 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc 5521 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc 5581 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga 5641 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt 5701 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg 5761 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa 5821 ggatggatgg ctacattctg gagacatagc ttactgggac gaggacgaac acttcttcat 5881 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt 5941 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtctgcccga 6001 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac 6061 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg 6121 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa 6181 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgg gttcggatta 6241 taaggaccat gacggagact ataaggacca tgacctcgac gctgcagcag cggattataa 6301 ggacgatgac gataagtgag cttgtcctgc tttaatgaga tatgcgagaa gcctatgatc 6361 gcatgatatt tgctttcaat tctgttgtgc acgttgtaaa aaacctgagc atgtgtagct 6421 cagatcctta ccgccggttt cggttcattc taatgaatat atcacccgtt actatcgtat 6481 ttttatgaat aatattctcc gttcaattta ctgattgtac cctactactt atatgtacaa 6541 tattaaaatg aaaacaatat attgtgctga ataggtttat agcgacatct atgatagagc 6601 gccacaataa caaacaattg cgttttatta ttacaaatcc aattttaaaa aaagcggcag 6661 aaccggtcaa acctaaaaga ctgattacat aaatcttatt caaatttcaa aagtgcccca 6721 ggggctagta tctacgacac accgagcggc gaactaataa cgctcactga agggaactcc 6781 ggttccccgc cggcgcgcat gggtgagatt ccttgaagtt gagtattggc cgtccgctct 6841 accgaaagtt acgggcacca ttcaacccgg tccagcacgg cggccgggta accgacttgc 6901 tgccccgaga attatgcagc atttttttgg tgtatgtggg ccccaaatga agtgcaggtc 6961 aaaccttgac agtgacgaca aatcgttggg cgggtccagg gcgaattttg cgacaacatg 7021 tcgaggctca gcaggaccgc t //