LOCUS Exported 7312 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo Level 1 luciferase reporter. ACCESSION . VERSION . KEYWORDS pEPYC1CB0321 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7312) 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 7312) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7312 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..632 /label=oriV /note="incP origin of replication" misc_feature 724..748 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" primer_bind 1202..1222 /label=35S promoter /note="CaMV 35S promoter, forward primer" promoter 1211..1257 /label=minimal CaMV 35S promoter /note="minimal 35S promoter from cauliflower mosaic virus" misc_feature 1278..1333 /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" CDS 1335..2984 /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(1401..1419) /label=LucNrev /note="Firefly luciferase, reverse primer" CDS 3048..3071 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" terminator 3082..3789 /label=OCS terminator /note="octopine synthase terminator" misc_feature 3838..3862 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS complement(4018..5166) /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(5502..6090) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5582..5601) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6261..7121) /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 6884..6903 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7122..7226) /gene="bla" /label=AmpR promoter ORIGIN 1 agcgggccgg gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc 61 acagggcgca gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa 121 agacaggtta gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc 181 ctgtggacag cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct 241 caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc 301 gcagggcact tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg 361 cgttttcgcc gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc 421 cctcatctgt caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc 481 atctgtcagt gagggccaag ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc 541 tcgcacacgg cttcgacggc gtttctggcg cgtttgcagg gccatagacg gccgccagcc 601 cagcggcgag ggcaaccagc ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg 661 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc 721 tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt 781 gcggacgttt ttaatgtact ggggtggatg cagtgggccc cactctgtga agacaatgcc 841 gaattcggat ccggaggtca acatggtgga gcacgacact ctggtctact ccaaaaatgt 901 caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa ggatgattcc 961 attgcccagc tatctgtcac ttcatcgaaa ggacagtaga aaaggaaggt ggctcctaca 1021 aatgccatca ttgcgataaa ggaaaggcta tcattcaaga tctctctgcc gacagtggtc 1081 ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagaggtt ccaaccacgt 1141 ctacaaagca agtggattga tgtgacatct ccactgacgt aagggatgac gcacaatccc 1201 actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac 1261 gctactgtat ttttacaaca attaccaaca acaacaaaca acaaacaaca ttacaattac 1321 tatttacaat tacaatggag gacgccaaaa acataaagaa aggcccggcg ccattctatc 1381 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg 1441 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt 1501 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc 1561 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt 1621 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca 1681 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa 1741 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa 1801 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt 1861 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca 1921 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct 1981 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg 2041 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt 2101 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagaactg tttctgagga 2161 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg 2221 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg 2281 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta 2341 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg 2401 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc 2461 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta 2521 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg 2581 gatggctaca ttctggagac atagcttact gggacgagga cgaacacttc ttcatcgttg 2641 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat 2701 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctg cccgacgatg 2761 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa 2821 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag 2881 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca 2941 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgggttcg gattataagg 3001 accatgacgg agactataag gaccatgacc tcgacgctgc agcagcggat tataaggacg 3061 atgacgataa gtgagcttgt cctgctttaa tgagatatgc gagaagccta tgatcgcatg 3121 atatttgctt tcaattctgt tgtgcacgtt gtaaaaaacc tgagcatgtg tagctcagat 3181 ccttaccgcc ggtttcggtt cattctaatg aatatatcac ccgttactat cgtattttta 3241 tgaataatat tctccgttca atttactgat tgtaccctac tacttatatg tacaatatta 3301 aaatgaaaac aatatattgt gctgaatagg tttatagcga catctatgat agagcgccac 3361 aataacaaac aattgcgttt tattattaca aatccaattt taaaaaaagc ggcagaaccg 3421 gtcaaaccta aaagactgat tacataaatc ttattcaaat ttcaaaagtg ccccaggggc 3481 tagtatctac gacacaccga gcggcgaact aataacgctc actgaaggga actccggttc 3541 cccgccggcg cgcatgggtg agattccttg aagttgagta ttggccgtcc gctctaccga 3601 aagttacggg caccattcaa cccggtccag cacggcggcc gggtaaccga cttgctgccc 3661 cgagaattat gcagcatttt tttggtgtat gtgggcccca aatgaagtgc aggtcaaacc 3721 ttgacagtga cgacaaatcg ttgggcgggt ccagggcgaa ttttgcgaca acatgtcgag 3781 gctcagcagg accgctgcaa ttgtcttctg cacgaagtgg tttaaactat cagtgtttga 3841 caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt 3901 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg 3961 ttccccagat caggcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta 4021 gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg 4081 caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga 4141 tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa 4201 atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt 4261 cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg 4321 ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag 4381 gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc 4441 ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta 4501 gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg 4561 ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc 4621 cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg 4681 cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca 4741 cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag 4801 caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg 4861 cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc 4921 ctcctgttca agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg 4981 agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac 5041 ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc 5101 ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg 5161 attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc 5221 ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc 5281 ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac 5341 gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca 5401 cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta 5461 ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 5521 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 5581 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 5641 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 5701 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5761 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5821 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5881 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5941 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 6001 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 6061 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 6121 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 6181 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6241 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6301 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6361 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct 6421 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6481 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6541 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6601 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6661 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6721 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6781 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6841 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6901 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6961 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7021 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7081 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 7141 tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag 7201 aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt tttggggtga 7261 ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg tt //