LOCUS Exported 8600 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MoClo Level 1 reporter. ACCESSION . VERSION . KEYWORDS pEPYC1CB0305 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8600) 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 8600) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8600 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 105..129 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" rep_origin 221..852 /label=oriV /note="incP origin of replication" promoter 939..1043 /gene="bla" /label=AmpR promoter CDS 1044..1904 /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(1262..1281) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2075..2663 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2564..2583 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2999..4147 /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 4303..4327 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 7159..7878 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(7204..7225) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(7465..7484) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7812..7833 /label=EGFP-C /note="EGFP, forward primer" terminator 7886..8593 /label=OCS terminator /note="octopine synthase terminator" ORIGIN 1 aagcttgaat tcttgcttgt cttcacagag tggggcccac tgcatccacc ccagtacatt 61 aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat 121 atcctgccac cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc 181 gatacaggca gcccatcagt cagatcagga tctcctttgc gacgctcacc gggctggttg 241 ccctcgccgc tgggctggcg gccgtctatg gccctgcaaa cgcgccagaa acgccgtcga 301 agccgtgtgc gagacaccgc ggccgccggc gttgtggata cctcgcggaa aacttggccc 361 tcactgacag atgaggggcg gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt 421 tgacagatga ggggcaggct cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa 481 tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat gatgtggaca agcctgggga 541 taagtgccct gcggtattga cacttgaggg gcgcgactac tgacagatga ggggcgcgat 601 ccttgacact tgaggggcag agtgctgaca gatgaggggc gcacctattg acatttgagg 661 ggctgtccac aggcagaaaa tccagcattt gcaagggttt ccgcccgttt ttcggccacc 721 gctaacctgt cttttaacct gcttttaaac caatatttat aaaccttgtt tttaaccagg 781 gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc 841 tcccggcccg ctaacgcggg cctcccatcc ccccaggggc tgcgcccctc ggccgcgaac 901 ggcctcaccc caaaaatggc agcgctggcc aattcgtgcg cggaacccct atttgtttat 961 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 1021 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct 1081 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag 1141 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta 1201 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc 1261 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca 1321 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg 1381 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg 1441 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca 1501 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa 1561 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa 1621 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata 1681 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat 1741 ctggagccgg tgagcgtggt tctcgcggta tcattgcagc actggggcca gatggtaagc 1801 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata 1861 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt 1921 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 1981 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 2041 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 2101 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 2161 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 2221 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 2281 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 2341 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 2401 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 2461 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 2521 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 2581 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 2641 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcag 2701 atcctagatg tggcgcaacg atgccggcga caagcaggag cgcaccgact tcttccgcat 2761 caagtgtttt ggctctcagg ccgaggccca cggcaagtat ttgggcaagg ggtcgctggt 2821 attcgtgcag ggcaagattc ggaataccaa gtacgagaag gacggccaga cggtctacgg 2881 gaccgacttc attgccgata aggtggatta tctggacacc aaggcaccag gcgggtcaaa 2941 tcaggaataa gggcacattg ccccggcgtg agtcggggca atcccgcaag gagggtgaat 3001 gaatcggacg tttgaccgga aggcatacag gcaagaactg atcgacgcgg ggttttccgc 3061 cgaggatgcc gaaaccatcg caagccgcac cgtcatgcgt gcgccccgcg aaaccttcca 3121 gtccgtcggc tcgatggtcc agcaagctac ggccaagatc gagcgcgaca gcgtgcaact 3181 ggctccccct gccctgcccg cgccatcggc cgccgtggag cgttcgcgtc gtcttgaaca 3241 ggaggcggca ggtttggcga agtcgatgac catcgacacg cgaggaacta tgacgaccaa 3301 gaagcgaaaa accgccggcg aggacctggc aaaacaggtc agcgaggcca agcaggccgc 3361 gttgctgaaa cacacgaagc agcagatcaa ggaaatgcag ctttccttgt tcgatattgc 3421 gccgtggccg gacacgatgc gagcgatgcc aaacgacacg gcccgctctg ccctgttcac 3481 cacgcgcaac aagaaaatcc cgcgcgaggc gctgcaaaac aaggtcattt tccacgtcaa 3541 caaggacgtg aagatcacct acaccggcgt cgagctgcgg gccgacgatg acgaactggt 3601 gtggcagcag gtgttggagt acgcgaagcg cacccctatc ggcgagccga tcaccttcac 3661 gttctacgag ctttgccagg acctgggctg gtcgatcaat ggccggtatt acacgaaggc 3721 cgaggaatgc ctgtcgcgcc tacaggcgac ggcgatgggc ttcacgtccg accgcgttgg 3781 gcacctggaa tcggtgtcgc tgctgcaccg cttccgcgtc ctggaccgtg gcaagaaaac 3841 gtcccgttgc caggtcctga tcgacgagga aatcgtcgtg ctgtttgctg gcgaccacta 3901 cacgaaattc atatgggaga agtaccgcaa gctgtcgccg acggcccgac ggatgttcga 3961 ctatttcagc tcgcaccggg agccgtaccc gctcaagctg gaaaccttcc gcctcatgtg 4021 cggatcggat tccacccgcg tgaagaagtg gcgcgagcag gtcggcgaag cctgcgaaga 4081 gttgcgaggc agcggcctgg tggaacacgc ctgggtcaat gatgacctgg tgcattgcaa 4141 acgctagggc cttgtggggt cagttccggc tgggggttca gcagccagcg cctgatctgg 4201 ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct 4261 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat 4321 cctgtcaaac actgatagtt taaaccactt cgtgcagaag acaatagtgg agggaggcgt 4381 gtcgttttag tgaggaagag gttccaacca cgtctacaaa accattattg cgcgttcaaa 4441 agctaaaaca ttgctttgtc aaaagctaaa aaagatgatg cccgacagcc agacgaagga 4501 aatgacgtaa gccatgacgt ctaatcccac aagctatata aggttttgct attcattgaa 4561 agcagtagtg actgatttgt atatatactg tattaaaatc ttaataggtt ttgataaaag 4621 cgaacgtggg gaaacccgaa ccaaaccttc ttctaaactc tctctcatct ctcttaaagc 4681 aaacttctct cttgtctttc ttgcgtgagc gatcttcaac gttgtcagat cgtgcttcgg 4741 caccagtaca acgttttctt tcactgaagc gaaatcaaag atctctttgt ggacacgtag 4801 tgcggcgcca ttaaataacg tgtacttgtc ctattcttgt cggtgtggtc ttgggaaaag 4861 aaagcttgct ggaggctgct gttcagcccc atacattact tgttacgatt ctgctgactt 4921 tcggcgggtg caatatctct acttctgctt gacgaggtat tgttgcctgt acttctttct 4981 tcttcttctt gctgattggt tctataagaa atctagtatt ttctttgaaa cagagttttc 5041 ccgtggtttt cgaacttgga gaaagattgt taagcttctg tatattctgc ccaaattacc 5101 atggtgagcg gctggcggct gttcaagaag attagcggaa tgggtcagtc ccttatgtta 5161 cgtcctgtag aaaccccaac ccgtgaaatc aaaaaactcg acggcctgtg ggcattcagt 5221 ctggatcgcg aaaactgtgg aattgatcag cgttggtggg aaagcgcgtt acaagaaagc 5281 cgggcaattg ctgtgccagg cagttttaac gatcagttcg ccgatgcaga tattcgtaat 5341 tatgcgggca acgtctggta tcagcgcgaa gtctttatac cgaaaggtaa gtcttactct 5401 ctcttttttg gtctgtattt ttaatttttt gaagtatact atttgtactg acgctaataa 5461 tcttttttca ggttgggcag gccagcgtat cgtgctgcgt ttcgatgcgg tcactcatta 5521 cggcaaagtg tgggtcaata atcaggaagt gatggagcat cagggcggct atacgccatt 5581 tgaagccgat gtcacgccgt atgttattgc cgggaaaagt gtacgtatca ccgtttgtgt 5641 gaacaacgaa ctgaactggc agactatccc gccgggaatg gtgattaccg acgaaaacgg 5701 caagaaaaag cagtcttact tccatgattt ctttaactat gccggaatcc atcgcagcgt 5761 aatgctctac accacgccga acacctgggt ggacgatatc accgtggtga cgcatgtcgc 5821 gcaagactgt aaccacgcgt ctgttgactg gcaggtactt catgcttcaa cgtgtaactt 5881 aagagatact gtgtgaaatt ttatatttcc atacatttgc ttgacctttg ctttttgtca 5941 atttttttcc ccttacaggt ggtggccaat ggtgatgtca gcgttgaact gcgtgatgcg 6001 gatcaacagg tggttgcaac tggacaaggc actagcggga ctttgcaagt ggtgaatccg 6061 cacctctggc aaccgggtga aggttatctc tatgaactgt gcgtcacagc caaaagccag 6121 acagagtgtg atatctaccc gcttcgcgtc ggcatccggt cagtggcagt gaagggcgaa 6181 cagttcctga ttaaccacaa accgttctac tttactggct ttggtcgtca tgaagatgcg 6241 gacttgcgtg gcaaaggatt cgataacgtg ctgatggtgc acgaccacgc attaatggac 6301 tggattgggg ccaactccta ccgtacctcg cattaccctt acgctgaaga gatgctcgac 6361 tgggcagatg aacatggcat cgtggtgatt gatgaaactg ctgctgtcgg ctttaacctc 6421 tctttaggca ttggtttcga agcgggcaac aagccgaaag aactgtacag cgaagaggca 6481 gtcaacgggg aaactcagca agcgcactta caggcgatta aagagctgat agcgcgtgac 6541 aaaaaccacc caagcgtggt gatgtggagt attgccaacg aaccggatac ccgtccgcaa 6601 ggtgcacggg aatatttcgc gccactggcg gaagcaacgc gtaaactcga cccgacgcgt 6661 ccgatcacct gcgtcaatgt aatgttctgc gacgctcaca ccgataccat cagcgatctc 6721 tttgatgtgc tgtgcctgaa ccgttattac ggatggtatg tccaaagcgg cgatttggaa 6781 acggcagaga aggtactgga aaaagaactt ctggcctggc aggagaaact gcatcagccg 6841 attatcatca ccgaatacgg cgtggatacg ttagccgggc tgcactcaat gtacaccgac 6901 atgtggagtg aagagtatca gtgtgcatgg ctggatatgt atcaccgcgt ctttgatcgc 6961 gtcagcgccg tcgtcggtga acaggtatgg aatttcgccg attttgcgac ctcgcaaggc 7021 atattgcgcg ttggcggtaa caagaaaggg atcttcactc gcgaccgcaa accgaagtcg 7081 gcggcttttc tgctgcaaaa acgctggact ggcatgaact tcggtgaaaa accgcagcag 7141 ggaggcaaac aaggttcgat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 7201 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 7261 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 7321 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc 7381 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 7441 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 7501 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 7561 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg 7621 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac 7681 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 7741 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag 7801 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 7861 gacgagctgt acaagtgagc ttgtcctgct ttaatgagat atgcgagaag cctatgatcg 7921 catgatattt gctttcaatt ctgttgtgca cgttgtaaaa aacctgagca tgtgtagctc 7981 agatccttac cgccggtttc ggttcattct aatgaatata tcacccgtta ctatcgtatt 8041 tttatgaata atattctccg ttcaatttac tgattgtacc ctactactta tatgtacaat 8101 attaaaatga aaacaatata ttgtgctgaa taggtttata gcgacatcta tgatagagcg 8161 ccacaataac aaacaattgc gttttattat tacaaatcca attttaaaaa aagcggcaga 8221 accggtcaaa cctaaaagac tgattacata aatcttattc aaatttcaaa agtgccccag 8281 gggctagtat ctacgacaca ccgagcggcg aactaataac gctcactgaa gggaactccg 8341 gttccccgcc ggcgcgcatg ggtgagattc cttgaagttg agtattggcc gtccgctcta 8401 ccgaaagtta cgggcaccat tcaacccggt ccagcacggc ggccgggtaa ccgacttgct 8461 gccccgagaa ttatgcagca tttttttggt gtatgtgggc cccaaatgaa gtgcaggtca 8521 aaccttgaca gtgacgacaa atcgttgggc gggtccaggg cgaattttgc gacaacatgt 8581 cgaggctcag caggaccgct //