LOCUS Exported 8663 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo Level 1 reporter. ACCESSION . VERSION . KEYWORDS pEPYC1CB0478 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8663) 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 8663) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8663 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 925..2073 /codon_start=1 /product="trans-acting replication protein that binds to and activates oriV" /label=trfA /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" misc_feature 2229..2253 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 5148..5867 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(5193..5214) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5454..5473) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5801..5822 /label=EGFP-C /note="EGFP, forward primer" terminator 5875..6582 /label=OCS terminator /note="octopine synthase terminator" misc_feature 6694..6718 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" rep_origin 6810..7441 /label=oriV /note="incP origin of replication" promoter 7528..7632 /gene="bla" /label=AmpR promoter CDS 7633..8493 /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 complement(7851..7870) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgta cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcggccttt ttacggttcc tggcagatcc tagatgtggc gcaacgatgc cggcgacaag 661 caggagcgca ccgacttctt ccgcatcaag tgttttggct ctcaggccga ggcccacggc 721 aagtatttgg gcaaggggtc gctggtattc gtgcagggca agattcggaa taccaagtac 781 gagaaggacg gccagacggt ctacgggacc gacttcattg ccgataaggt ggattatctg 841 gacaccaagg caccaggcgg gtcaaatcag gaataagggc acattgcccc ggcgtgagtc 901 ggggcaatcc cgcaaggagg gtgaatgaat cggacgtttg accggaaggc atacaggcaa 961 gaactgatcg acgcggggtt ttccgccgag gatgccgaaa ccatcgcaag ccgcaccgtc 1021 atgcgtgcgc cccgcgaaac cttccagtcc gtcggctcga tggtccagca agctacggcc 1081 aagatcgagc gcgacagcgt gcaactggct ccccctgccc tgcccgcgcc atcggccgcc 1141 gtggagcgtt cgcgtcgtct tgaacaggag gcggcaggtt tggcgaagtc gatgaccatc 1201 gacacgcgag gaactatgac gaccaagaag cgaaaaaccg ccggcgagga cctggcaaaa 1261 caggtcagcg aggccaagca ggccgcgttg ctgaaacaca cgaagcagca gatcaaggaa 1321 atgcagcttt ccttgttcga tattgcgccg tggccggaca cgatgcgagc gatgccaaac 1381 gacacggccc gctctgccct gttcaccacg cgcaacaaga aaatcccgcg cgaggcgctg 1441 caaaacaagg tcattttcca cgtcaacaag gacgtgaaga tcacctacac cggcgtcgag 1501 ctgcgggccg acgatgacga actggtgtgg cagcaggtgt tggagtacgc gaagcgcacc 1561 cctatcggcg agccgatcac cttcacgttc tacgagcttt gccaggacct gggctggtcg 1621 atcaatggcc ggtattacac gaaggccgag gaatgcctgt cgcgcctaca ggcgacggcg 1681 atgggcttca cgtccgaccg cgttgggcac ctggaatcgg tgtcgctgct gcaccgcttc 1741 cgcgtcctgg accgtggcaa gaaaacgtcc cgttgccagg tcctgatcga cgaggaaatc 1801 gtcgtgctgt ttgctggcga ccactacacg aaattcatat gggagaagta ccgcaagctg 1861 tcgccgacgg cccgacggat gttcgactat ttcagctcgc accgggagcc gtacccgctc 1921 aagctggaaa ccttccgcct catgtgcgga tcggattcca cccgcgtgaa gaagtggcgc 1981 gagcaggtcg gcgaagcctg cgaagagttg cgaggcagcg gcctggtgga acacgcctgg 2041 gtcaatgatg acctggtgca ttgcaaacgc tagggccttg tggggtcagt tccggctggg 2101 ggttcagcag ccagcgcctg atctggggaa ccctgtggtt ggcatgcaca tacaaatgga 2161 cgaacggata aaccttttca cgccctttta aatatccgat tattctaata aacgctcttt 2221 tctcttaggt ttacccgcca atatatcctg tcaaacactg atagtttaaa ccacttcgtg 2281 cagaagacaa tagtggaggg aggcgtgtcg ttttagtgag gagcaagtgg atgtaggtca 2341 cgaccatcac tatcagcgat cttgcctgcc ttttatgacc cccgccgatg acgcgggaag 2401 gaggaccgat gctgattctc tctgccgaca gtggtcccaa aggttgtctg caccaaccat 2461 tattgcgtca catgtaggct atcagcttat gacgtaagcc atgacgtcta gacctctaca 2521 aactatataa ggttttgcta ttcattgaaa gcagtagtga ctgatttgta tatatactgt 2581 attaaaatct taataggttt tgataaaagc gaacgtgggg aaacccgaac caaaccttct 2641 tctaaactct ctctcatctc tcttaaagca aacttctctc ttgtctttct tgcgtgagcg 2701 atcttcaacg ttgtcagatc gtgcttcggc accagtacaa cgttttcttt cactgaagcg 2761 aaatcaaaga tctctttgtg gacacgtagt gcggcgccat taaataacgt gtacttgtcc 2821 tattcttgtc ggtgtggtct tgggaaaaga aagcttgctg gaggctgctg ttcagcccca 2881 tacattactt gttacgattc tgctgacttt cggcgggtgc aatatctcta cttctgcttg 2941 acgaggtatt gttgcctgta cttctttctt cttcttcttg ctgattggtt ctataagaaa 3001 tctagtattt tctttgaaac agagttttcc cgtggttttc gaacttggag aaagattgtt 3061 aagcttctgt atattctgcc caaattacca tggtgagcgg ctggcggctg ttcaagaaga 3121 ttagcggaat gggtcagtcc cttatgttac gtcctgtaga aaccccaacc cgtgaaatca 3181 aaaaactcga cggcctgtgg gcattcagtc tggatcgcga aaactgtgga attgatcagc 3241 gttggtggga aagcgcgtta caagaaagcc gggcaattgc tgtgccaggc agttttaacg 3301 atcagttcgc cgatgcagat attcgtaatt atgcgggcaa cgtctggtat cagcgcgaag 3361 tctttatacc gaaaggtaag tcttactctc tcttttttgg tctgtatttt taattttttg 3421 aagtatacta tttgtactga cgctaataat cttttttcag gttgggcagg ccagcgtatc 3481 gtgctgcgtt tcgatgcggt cactcattac ggcaaagtgt gggtcaataa tcaggaagtg 3541 atggagcatc agggcggcta tacgccattt gaagccgatg tcacgccgta tgttattgcc 3601 gggaaaagtg tacgtatcac cgtttgtgtg aacaacgaac tgaactggca gactatcccg 3661 ccgggaatgg tgattaccga cgaaaacggc aagaaaaagc agtcttactt ccatgatttc 3721 tttaactatg ccggaatcca tcgcagcgta atgctctaca ccacgccgaa cacctgggtg 3781 gacgatatca ccgtggtgac gcatgtcgcg caagactgta accacgcgtc tgttgactgg 3841 caggtacttc atgcttcaac gtgtaactta agagatactg tgtgaaattt tatatttcca 3901 tacatttgct tgacctttgc tttttgtcaa tttttttccc cttacaggtg gtggccaatg 3961 gtgatgtcag cgttgaactg cgtgatgcgg atcaacaggt ggttgcaact ggacaaggca 4021 ctagcgggac tttgcaagtg gtgaatccgc acctctggca accgggtgaa ggttatctct 4081 atgaactgtg cgtcacagcc aaaagccaga cagagtgtga tatctacccg cttcgcgtcg 4141 gcatccggtc agtggcagtg aagggcgaac agttcctgat taaccacaaa ccgttctact 4201 ttactggctt tggtcgtcat gaagatgcgg acttgcgtgg caaaggattc gataacgtgc 4261 tgatggtgca cgaccacgca ttaatggact ggattggggc caactcctac cgtacctcgc 4321 attaccctta cgctgaagag atgctcgact gggcagatga acatggcatc gtggtgattg 4381 atgaaactgc tgctgtcggc tttaacctct ctttaggcat tggtttcgaa gcgggcaaca 4441 agccgaaaga actgtacagc gaagaggcag tcaacgggga aactcagcaa gcgcacttac 4501 aggcgattaa agagctgata gcgcgtgaca aaaaccaccc aagcgtggtg atgtggagta 4561 ttgccaacga accggatacc cgtccgcaag gtgcacggga atatttcgcg ccactggcgg 4621 aagcaacgcg taaactcgac ccgacgcgtc cgatcacctg cgtcaatgta atgttctgcg 4681 acgctcacac cgataccatc agcgatctct ttgatgtgct gtgcctgaac cgttattacg 4741 gatggtatgt ccaaagcggc gatttggaaa cggcagagaa ggtactggaa aaagaacttc 4801 tggcctggca ggagaaactg catcagccga ttatcatcac cgaatacggc gtggatacgt 4861 tagccgggct gcactcaatg tacaccgaca tgtggagtga agagtatcag tgtgcatggc 4921 tggatatgta tcaccgcgtc tttgatcgcg tcagcgccgt cgtcggtgaa caggtatgga 4981 atttcgccga ttttgcgacc tcgcaaggca tattgcgcgt tggcggtaac aagaaaggga 5041 tcttcactcg cgaccgcaaa ccgaagtcgg cggcttttct gctgcaaaaa cgctggactg 5101 gcatgaactt cggtgaaaaa ccgcagcagg gaggcaaaca aggttcgatg gtgagcaagg 5161 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg 5221 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc 5281 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct 5341 tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct 5401 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg 5461 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg 5521 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca 5581 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga 5641 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc 5701 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc 5761 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg 5821 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgagct tgtcctgctt 5881 taatgagata tgcgagaagc ctatgatcgc atgatatttg ctttcaattc tgttgtgcac 5941 gttgtaaaaa acctgagcat gtgtagctca gatccttacc gccggtttcg gttcattcta 6001 atgaatatat cacccgttac tatcgtattt ttatgaataa tattctccgt tcaatttact 6061 gattgtaccc tactacttat atgtacaata ttaaaatgaa aacaatatat tgtgctgaat 6121 aggtttatag cgacatctat gatagagcgc cacaataaca aacaattgcg ttttattatt 6181 acaaatccaa ttttaaaaaa agcggcagaa ccggtcaaac ctaaaagact gattacataa 6241 atcttattca aatttcaaaa gtgccccagg ggctagtatc tacgacacac cgagcggcga 6301 actaataacg ctcactgaag ggaactccgg ttccccgccg gcgcgcatgg gtgagattcc 6361 ttgaagttga gtattggccg tccgctctac cgaaagttac gggcaccatt caacccggtc 6421 cagcacggcg gccgggtaac cgacttgctg ccccgagaat tatgcagcat ttttttggtg 6481 tatgtgggcc ccaaatgaag tgcaggtcaa accttgacag tgacgacaaa tcgttgggcg 6541 ggtccagggc gaattttgcg acaacatgtc gaggctcagc aggaccgcta agcttgaatt 6601 cttgcttgtc ttcacagagt ggggcccact gcatccaccc cagtacatta aaaacgtccg 6661 caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc 6721 agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag 6781 cccatcagtc agatcaggat ctcctttgcg acgctcaccg ggctggttgc cctcgccgct 6841 gggctggcgg ccgtctatgg ccctgcaaac gcgccagaaa cgccgtcgaa gccgtgtgcg 6901 agacaccgcg gccgccggcg ttgtggatac ctcgcggaaa acttggccct cactgacaga 6961 tgaggggcgg acgttgacac ttgaggggcc gactcacccg gcgcggcgtt gacagatgag 7021 gggcaggctc gatttcggcc ggcgacgtgg agctggccag cctcgcaaat cggcgaaaac 7081 gcctgatttt acgcgagttt cccacagatg atgtggacaa gcctggggat aagtgccctg 7141 cggtattgac acttgagggg cgcgactact gacagatgag gggcgcgatc cttgacactt 7201 gaggggcaga gtgctgacag atgaggggcg cacctattga catttgaggg gctgtccaca 7261 ggcagaaaat ccagcatttg caagggtttc cgcccgtttt tcggccaccg ctaacctgtc 7321 ttttaacctg cttttaaacc aatatttata aaccttgttt ttaaccaggg ctgcgccctg 7381 tgcgcgtgac cgcgcacgcc gaaggggggt gccccccctt ctcgaaccct cccggcccgc 7441 taacgcgggc ctcccatccc cccaggggct gcgcccctcg gccgcgaacg gcctcacccc 7501 aaaaatggca gcgctggcca attcgtgcgc ggaaccccta tttgtttatt tttctaaata 7561 cattcaaata tgtatccgct catgagacaa taaccttgat aaatgcttca ataatattga 7621 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca 7681 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat 7741 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag 7801 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc 7861 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct 7921 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca 7981 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt 8041 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat 8101 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt 8161 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta 8221 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga 8281 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt 8341 gagcgtggtt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc 8401 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct 8461 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata 8521 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 8581 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 8641 gtagaaaaga tcaaaggatc ttc //