LOCUS Exported 7056 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo Level 1 luciferase reporter. ACCESSION . VERSION . KEYWORDS pEPYC1CB0442 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7056) 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 7056) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7056 /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 4538..4593 /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" CDS 4595..6244 /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(4661..4679) /label=LucNrev /note="Firefly luciferase, reverse primer" CDS 6308..6331 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 6342..7049 /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 tgaggatcct taccgctatg ggtaagatta tccttaccgc tatgggtaag 4441 attatcctta ccgctatggg taagattcta tataaggttt tgctattcat tgaaagcagt 4501 agtgactgat ttgtatataa tgtactgtat ttttacaaca attaccaaca acaacaaaca 4561 acaaacaaca ttacaattac tatttacaat tacaatggag gacgccaaaa acataaagaa 4621 aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc 4681 tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt 4741 ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg 4801 atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt 4861 tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta 4921 taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc 4981 caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat 5041 tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac 5101 atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga 5161 caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc 5221 tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa 5281 tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat 5341 gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga 5401 agaagaactg tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc 5461 aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt 5521 acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa 5581 gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat 5641 tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt 5701 tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga 5761 actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa 5821 cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact gggacgagga 5881 cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt 5941 ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt 6001 cgcaggtctg cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca 6061 cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc 6121 gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa 6181 actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc 6241 cgtgggttcg gattataagg accatgacgg agactataag gaccatgacc tcgacgctgc 6301 agcagcggat tataaggacg atgacgataa gtgagcttgt cctgctttaa tgagatatgc 6361 gagaagccta tgatcgcatg atatttgctt tcaattctgt tgtgcacgtt gtaaaaaacc 6421 tgagcatgtg tagctcagat ccttaccgcc ggtttcggtt cattctaatg aatatatcac 6481 ccgttactat cgtattttta tgaataatat tctccgttca atttactgat tgtaccctac 6541 tacttatatg tacaatatta aaatgaaaac aatatattgt gctgaatagg tttatagcga 6601 catctatgat agagcgccac aataacaaac aattgcgttt tattattaca aatccaattt 6661 taaaaaaagc ggcagaaccg gtcaaaccta aaagactgat tacataaatc ttattcaaat 6721 ttcaaaagtg ccccaggggc tagtatctac gacacaccga gcggcgaact aataacgctc 6781 actgaaggga actccggttc cccgccggcg cgcatgggtg agattccttg aagttgagta 6841 ttggccgtcc gctctaccga aagttacggg caccattcaa cccggtccag cacggcggcc 6901 gggtaaccga cttgctgccc cgagaattat gcagcatttt tttggtgtat gtgggcccca 6961 aatgaagtgc aggtcaaacc ttgacagtga cgacaaatcg ttgggcgggt ccagggcgaa 7021 ttttgcgaca acatgtcgag gctcagcagg accgct //