LOCUS       sequence-318576-        8484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1 reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8484)
  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 8484)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2020 Aug 28. pii: 5897334. doi: 10.1093/nar/gkaa682."
COMMENT     Sequence Label: pEPYC1CB0503
FEATURES             Location/Qualifiers
     source          1..8484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             join(2741..2743,2744..2746,2747..3460)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2786..2807)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3047..3066)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3394..3415
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3468..4175
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    4287..4311
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      4403..5034
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5121..5225
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5226..5294,5295..6086)
                     /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(5444..5463)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6257..6845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6746..6765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7181..8329
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aaccacttcg tgcagaagac
       61 aatagtggag ggaggcgtgt cgttttagtg aggtccccgc atagctgaac atctatataa
      121 ggttttgcta ttcattgaaa gcagtagtga ctgatttgta tataatgtac tgtattaaaa
      181 tcttaatagg ttttgataaa agcgaacgtg gggaaacccg aaccaaacct tcttctaaac
      241 tctctctcat ctctcttaaa gcaaacttct ctcttgtctt tcttgcgtga gcgatcttca
      301 acgttgtcag atcgtgcttc ggcaccagta caacgttttc tttcactgaa gcgaaatcaa
      361 agatctcttt gtggacacgt agtgcggcgc cattaaataa cgtgtacttg tcctattctt
      421 gtcggtgtgg tcttgggaaa agaaagcttg ctggaggctg ctgttcagcc ccatacatta
      481 cttgttacga ttctgctgac tttcggcggg tgcaatatct ctacttctgc ttgacgaggt
      541 attgttgcct gtacttcttt cttcttcttc ttgctgattg gttctataag aaatctagta
      601 ttttctttga aacagagttt tcccgtggtt ttcgaacttg gagaaagatt gttaagcttc
      661 tgtatattct gcccaaatta ccatggtgag cggctggcgg ctgttcaaga agattagcgg
      721 aatgggtcag tcccttatgt tacgtcctgt agaaacccca acccgtgaaa tcaaaaaact
      781 cgacggcctg tgggcattca gtctggatcg cgaaaactgt ggaattgatc agcgttggtg
      841 ggaaagcgcg ttacaagaaa gccgggcaat tgctgtgcca ggcagtttta acgatcagtt
      901 cgccgatgca gatattcgta attatgcggg caacgtctgg tatcagcgcg aagtctttat
      961 accgaaaggt aagtcttact ctctcttttt tggtctgtat ttttaatttt ttgaagtata
     1021 ctatttgtac tgacgctaat aatctttttt caggttgggc aggccagcgt atcgtgctgc
     1081 gtttcgatgc ggtcactcat tacggcaaag tgtgggtcaa taatcaggaa gtgatggagc
     1141 atcagggcgg ctatacgcca tttgaagccg atgtcacgcc gtatgttatt gccgggaaaa
     1201 gtgtacgtat caccgtttgt gtgaacaacg aactgaactg gcagactatc ccgccgggaa
     1261 tggtgattac cgacgaaaac ggcaagaaaa agcagtctta cttccatgat ttctttaact
     1321 atgccggaat ccatcgcagc gtaatgctct acaccacgcc gaacacctgg gtggacgata
     1381 tcaccgtggt gacgcatgtc gcgcaagact gtaaccacgc gtctgttgac tggcaggtac
     1441 ttcatgcttc aacgtgtaac ttaagagata ctgtgtgaaa ttttatattt ccatacattt
     1501 gcttgacctt tgctttttgt caattttttt ccccttacag gtggtggcca atggtgatgt
     1561 cagcgttgaa ctgcgtgatg cggatcaaca ggtggttgca actggacaag gcactagcgg
     1621 gactttgcaa gtggtgaatc cgcacctctg gcaaccgggt gaaggttatc tctatgaact
     1681 gtgcgtcaca gccaaaagcc agacagagtg tgatatctac ccgcttcgcg tcggcatccg
     1741 gtcagtggca gtgaagggcg aacagttcct gattaaccac aaaccgttct actttactgg
     1801 ctttggtcgt catgaagatg cggacttgcg tggcaaagga ttcgataacg tgctgatggt
     1861 gcacgaccac gcattaatgg actggattgg ggccaactcc taccgtacct cgcattaccc
     1921 ttacgctgaa gagatgctcg actgggcaga tgaacatggc atcgtggtga ttgatgaaac
     1981 tgctgctgtc ggctttaacc tctctttagg cattggtttc gaagcgggca acaagccgaa
     2041 agaactgtac agcgaagagg cagtcaacgg ggaaactcag caagcgcact tacaggcgat
     2101 taaagagctg atagcgcgtg acaaaaacca cccaagcgtg gtgatgtgga gtattgccaa
     2161 cgaaccggat acccgtccgc aaggtgcacg ggaatatttc gcgccactgg cggaagcaac
     2221 gcgtaaactc gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca
     2281 caccgatacc atcagcgatc tctttgatgt gctgtgcctg aaccgttatt acggatggta
     2341 tgtccaaagc ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac ttctggcctg
     2401 gcaggagaaa ctgcatcagc cgattatcat caccgaatac ggcgtggata cgttagccgg
     2461 gctgcactca atgtacaccg acatgtggag tgaagagtat cagtgtgcat ggctggatat
     2521 gtatcaccgc gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc
     2581 cgattttgcg acctcgcaag gcatattgcg cgttggcggt aacaagaaag ggatcttcac
     2641 tcgcgaccgc aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa
     2701 cttcggtgaa aaaccgcagc agggaggcaa acaaggttcg atggtgagca agggcgagga
     2761 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2821 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2881 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
     2941 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3001 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3061 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3121 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3181 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     3241 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3301 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     3361 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3421 cgccgggatc actctcggca tggacgagct gtacaagtga gcttgtcctg ctttaatgag
     3481 atatgcgaga agcctatgat cgcatgatat ttgctttcaa ttctgttgtg cacgttgtaa
     3541 aaaacctgag catgtgtagc tcagatcctt accgccggtt tcggttcatt ctaatgaata
     3601 tatcacccgt tactatcgta tttttatgaa taatattctc cgttcaattt actgattgta
     3661 ccctactact tatatgtaca atattaaaat gaaaacaata tattgtgctg aataggttta
     3721 tagcgacatc tatgatagag cgccacaata acaaacaatt gcgttttatt attacaaatc
     3781 caattttaaa aaaagcggca gaaccggtca aacctaaaag actgattaca taaatcttat
     3841 tcaaatttca aaagtgcccc aggggctagt atctacgaca caccgagcgg cgaactaata
     3901 acgctcactg aagggaactc cggttccccg ccggcgcgca tgggtgagat tccttgaagt
     3961 tgagtattgg ccgtccgctc taccgaaagt tacgggcacc attcaacccg gtccagcacg
     4021 gcggccgggt aaccgacttg ctgccccgag aattatgcag catttttttg gtgtatgtgg
     4081 gccccaaatg aagtgcaggt caaaccttga cagtgacgac aaatcgttgg gcgggtccag
     4141 ggcgaatttt gcgacaacat gtcgaggctc agcaggaccg ctaagcttga attcttgctt
     4201 gtcttcacag agtggggccc actgcatcca ccccagtaca ttaaaaacgt ccgcaatgtg
     4261 ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc
     4321 caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca
     4381 gtcagatcag gatctccttt gcgacgctca ccgggctggt tgccctcgcc gctgggctgg
     4441 cggccgtcta tggccctgca aacgcgccag aaacgccgtc gaagccgtgt gcgagacacc
     4501 gcggccgccg gcgttgtgga tacctcgcgg aaaacttggc cctcactgac agatgagggg
     4561 cggacgttga cacttgaggg gccgactcac ccggcgcggc gttgacagat gaggggcagg
     4621 ctcgatttcg gccggcgacg tggagctggc cagcctcgca aatcggcgaa aacgcctgat
     4681 tttacgcgag tttcccacag atgatgtgga caagcctggg gataagtgcc ctgcggtatt
     4741 gacacttgag gggcgcgact actgacagat gaggggcgcg atccttgaca cttgaggggc
     4801 agagtgctga cagatgaggg gcgcacctat tgacatttga ggggctgtcc acaggcagaa
     4861 aatccagcat ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct gtcttttaac
     4921 ctgcttttaa accaatattt ataaaccttg tttttaacca gggctgcgcc ctgtgcgcgt
     4981 gaccgcgcac gccgaagggg ggtgcccccc cttctcgaac cctcccggcc cgctaacgcg
     5041 ggcctcccat ccccccaggg gctgcgcccc tcggccgcga acggcctcac cccaaaaatg
     5101 gcagcgctgg ccaattcgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5161 atatgtatcc gctcatgaga caataacctt gataaatgct tcaataatat tgaaaaagga
     5221 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5281 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5341 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5401 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5461 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5521 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5581 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5641 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5701 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5761 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5821 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5881 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5941 gttctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6001 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6061 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6121 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6181 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc agatcctaga tgtggcgcaa
     6901 cgatgccggc gacaagcagg agcgcaccga cttcttccgc atcaagtgtt ttggctctca
     6961 ggccgaggcc cacggcaagt atttgggcaa ggggtcgctg gtattcgtgc agggcaagat
     7021 tcggaatacc aagtacgaga aggacggcca gacggtctac gggaccgact tcattgccga
     7081 taaggtggat tatctggaca ccaaggcacc aggcgggtca aatcaggaat aagggcacat
     7141 tgccccggcg tgagtcgggg caatcccgca aggagggtga atgaatcgga cgtttgaccg
     7201 gaaggcatac aggcaagaac tgatcgacgc ggggttttcc gccgaggatg ccgaaaccat
     7261 cgcaagccgc accgtcatgc gtgcgccccg cgaaaccttc cagtccgtcg gctcgatggt
     7321 ccagcaagct acggccaaga tcgagcgcga cagcgtgcaa ctggctcccc ctgccctgcc
     7381 cgcgccatcg gccgccgtgg agcgttcgcg tcgtcttgaa caggaggcgg caggtttggc
     7441 gaagtcgatg accatcgaca cgcgaggaac tatgacgacc aagaagcgaa aaaccgccgg
     7501 cgaggacctg gcaaaacagg tcagcgaggc caagcaggcc gcgttgctga aacacacgaa
     7561 gcagcagatc aaggaaatgc agctttcctt gttcgatatt gcgccgtggc cggacacgat
     7621 gcgagcgatg ccaaacgaca cggcccgctc tgccctgttc accacgcgca acaagaaaat
     7681 cccgcgcgag gcgctgcaaa acaaggtcat tttccacgtc aacaaggacg tgaagatcac
     7741 ctacaccggc gtcgagctgc gggccgacga tgacgaactg gtgtggcagc aggtgttgga
     7801 gtacgcgaag cgcaccccta tcggcgagcc gatcaccttc acgttctacg agctttgcca
     7861 ggacctgggc tggtcgatca atggccggta ttacacgaag gccgaggaat gcctgtcgcg
     7921 cctacaggcg acggcgatgg gcttcacgtc cgaccgcgtt gggcacctgg aatcggtgtc
     7981 gctgctgcac cgcttccgcg tcctggaccg tggcaagaaa acgtcccgtt gccaggtcct
     8041 gatcgacgag gaaatcgtcg tgctgtttgc tggcgaccac tacacgaaat tcatatggga
     8101 gaagtaccgc aagctgtcgc cgacggcccg acggatgttc gactatttca gctcgcaccg
     8161 ggagccgtac ccgctcaagc tggaaacctt ccgcctcatg tgcggatcgg attccacccg
     8221 cgtgaagaag tggcgcgagc aggtcggcga agcctgcgaa gagttgcgag gcagcggcct
     8281 ggtggaacac gcctgggtca atgatgacct ggtgcattgc aaacgctagg gccttgtggg
     8341 gtcagttccg gctgggggtt cagcagccag cgcctgatct ggggaaccct gtggttggca
     8401 tgcacataca aatggacgaa cggataaacc ttttcacgcc cttttaaata tccgattatt
     8461 ctaataaacg ctcttttctc ttag
//
