LOCUS       sequence-318573-        8808 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 8808)
  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 8808)
  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: pEPYC1CB0477
FEATURES             Location/Qualifiers
     source          1..8808
                     /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(3065..3067,3068..3070,3071..3784)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3110..3131)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3371..3390)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3718..3739
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3792..4499
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    4611..4635
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      4727..5358
                     /label=oriV
                     /note="incP origin of replication"
     promoter        5445..5549
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5550..5618,5619..6410)
                     /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(5768..5787)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6581..7169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7070..7089
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7505..8653
                     /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 aggatcgaaa ggacagtaac tgctaggagg
      121 accgatgctg atcttccatc aacaaataat ccaagtaagg taggtcacga ccactataat
      181 ttcgggaaac ctcctcggct atcgctttag caagacctct acattgcgat aaaggaaagg
      241 ctatgcccaa ttatgacccc cgccgatgac gcgggaaaaa ctggtacttg ttgaagcatc
      301 ttccactatg cccaatggtg gagcacgaca aaactggtac ttgtgtacag ctgacgtaag
      361 ggatgacgca catatcagct tagcaagacc tcttgaagat aagataataa tgttgaagat
      421 aagatgcctg ccttgatgac tatataaggt tttgctattc attgaaagca gtagtgactg
      481 atttgtatat atactgtatt aaaatcttaa taggttttga taaaagcgaa cgtggggaaa
      541 cccgaaccaa accttcttct aaactctctc tcatctctct taaagcaaac ttctctcttg
      601 tctttcttgc gtgagcgatc ttcaacgttg tcagatcgtg cttcggcacc agtacaacgt
      661 tttctttcac tgaagcgaaa tcaaagatct ctttgtggac acgtagtgcg gcgccattaa
      721 ataacgtgta cttgtcctat tcttgtcggt gtggtcttgg gaaaagaaag cttgctggag
      781 gctgctgttc agccccatac attacttgtt acgattctgc tgactttcgg cgggtgcaat
      841 atctctactt ctgcttgacg aggtattgtt gcctgtactt ctttcttctt cttcttgctg
      901 attggttcta taagaaatct agtattttct ttgaaacaga gttttcccgt ggttttcgaa
      961 cttggagaaa gattgttaag cttctgtata ttctgcccaa attaccatgg tgagcggctg
     1021 gcggctgttc aagaagatta gcggaatggg tcagtccctt atgttacgtc ctgtagaaac
     1081 cccaacccgt gaaatcaaaa aactcgacgg cctgtgggca ttcagtctgg atcgcgaaaa
     1141 ctgtggaatt gatcagcgtt ggtgggaaag cgcgttacaa gaaagccggg caattgctgt
     1201 gccaggcagt tttaacgatc agttcgccga tgcagatatt cgtaattatg cgggcaacgt
     1261 ctggtatcag cgcgaagtct ttataccgaa aggtaagtct tactctctct tttttggtct
     1321 gtatttttaa ttttttgaag tatactattt gtactgacgc taataatctt ttttcaggtt
     1381 gggcaggcca gcgtatcgtg ctgcgtttcg atgcggtcac tcattacggc aaagtgtggg
     1441 tcaataatca ggaagtgatg gagcatcagg gcggctatac gccatttgaa gccgatgtca
     1501 cgccgtatgt tattgccggg aaaagtgtac gtatcaccgt ttgtgtgaac aacgaactga
     1561 actggcagac tatcccgccg ggaatggtga ttaccgacga aaacggcaag aaaaagcagt
     1621 cttacttcca tgatttcttt aactatgccg gaatccatcg cagcgtaatg ctctacacca
     1681 cgccgaacac ctgggtggac gatatcaccg tggtgacgca tgtcgcgcaa gactgtaacc
     1741 acgcgtctgt tgactggcag gtacttcatg cttcaacgtg taacttaaga gatactgtgt
     1801 gaaattttat atttccatac atttgcttga cctttgcttt ttgtcaattt ttttcccctt
     1861 acaggtggtg gccaatggtg atgtcagcgt tgaactgcgt gatgcggatc aacaggtggt
     1921 tgcaactgga caaggcacta gcgggacttt gcaagtggtg aatccgcacc tctggcaacc
     1981 gggtgaaggt tatctctatg aactgtgcgt cacagccaaa agccagacag agtgtgatat
     2041 ctacccgctt cgcgtcggca tccggtcagt ggcagtgaag ggcgaacagt tcctgattaa
     2101 ccacaaaccg ttctacttta ctggctttgg tcgtcatgaa gatgcggact tgcgtggcaa
     2161 aggattcgat aacgtgctga tggtgcacga ccacgcatta atggactgga ttggggccaa
     2221 ctcctaccgt acctcgcatt acccttacgc tgaagagatg ctcgactggg cagatgaaca
     2281 tggcatcgtg gtgattgatg aaactgctgc tgtcggcttt aacctctctt taggcattgg
     2341 tttcgaagcg ggcaacaagc cgaaagaact gtacagcgaa gaggcagtca acggggaaac
     2401 tcagcaagcg cacttacagg cgattaaaga gctgatagcg cgtgacaaaa accacccaag
     2461 cgtggtgatg tggagtattg ccaacgaacc ggatacccgt ccgcaaggtg cacgggaata
     2521 tttcgcgcca ctggcggaag caacgcgtaa actcgacccg acgcgtccga tcacctgcgt
     2581 caatgtaatg ttctgcgacg ctcacaccga taccatcagc gatctctttg atgtgctgtg
     2641 cctgaaccgt tattacggat ggtatgtcca aagcggcgat ttggaaacgg cagagaaggt
     2701 actggaaaaa gaacttctgg cctggcagga gaaactgcat cagccgatta tcatcaccga
     2761 atacggcgtg gatacgttag ccgggctgca ctcaatgtac accgacatgt ggagtgaaga
     2821 gtatcagtgt gcatggctgg atatgtatca ccgcgtcttt gatcgcgtca gcgccgtcgt
     2881 cggtgaacag gtatggaatt tcgccgattt tgcgacctcg caaggcatat tgcgcgttgg
     2941 cggtaacaag aaagggatct tcactcgcga ccgcaaaccg aagtcggcgg cttttctgct
     3001 gcaaaaacgc tggactggca tgaacttcgg tgaaaaaccg cagcagggag gcaaacaagg
     3061 ttcgatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     3121 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     3181 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     3241 caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat
     3301 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3361 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3421 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3481 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3541 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3601 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3661 ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3721 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3781 gtgagcttgt cctgctttaa tgagatatgc gagaagccta tgatcgcatg atatttgctt
     3841 tcaattctgt tgtgcacgtt gtaaaaaacc tgagcatgtg tagctcagat ccttaccgcc
     3901 ggtttcggtt cattctaatg aatatatcac ccgttactat cgtattttta tgaataatat
     3961 tctccgttca atttactgat tgtaccctac tacttatatg tacaatatta aaatgaaaac
     4021 aatatattgt gctgaatagg tttatagcga catctatgat agagcgccac aataacaaac
     4081 aattgcgttt tattattaca aatccaattt taaaaaaagc ggcagaaccg gtcaaaccta
     4141 aaagactgat tacataaatc ttattcaaat ttcaaaagtg ccccaggggc tagtatctac
     4201 gacacaccga gcggcgaact aataacgctc actgaaggga actccggttc cccgccggcg
     4261 cgcatgggtg agattccttg aagttgagta ttggccgtcc gctctaccga aagttacggg
     4321 caccattcaa cccggtccag cacggcggcc gggtaaccga cttgctgccc cgagaattat
     4381 gcagcatttt tttggtgtat gtgggcccca aatgaagtgc aggtcaaacc ttgacagtga
     4441 cgacaaatcg ttgggcgggt ccagggcgaa ttttgcgaca acatgtcgag gctcagcagg
     4501 accgctaagc ttgaattctt gcttgtcttc acagagtggg gcccactgca tccaccccag
     4561 tacattaaaa acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca
     4621 caatatatcc tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca
     4681 ccactcgata caggcagccc atcagtcaga tcaggatctc ctttgcgacg ctcaccgggc
     4741 tggttgccct cgccgctggg ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc
     4801 cgtcgaagcc gtgtgcgaga caccgcggcc gccggcgttg tggatacctc gcggaaaact
     4861 tggccctcac tgacagatga ggggcggacg ttgacacttg aggggccgac tcacccggcg
     4921 cggcgttgac agatgagggg caggctcgat ttcggccggc gacgtggagc tggccagcct
     4981 cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc acagatgatg tggacaagcc
     5041 tggggataag tgccctgcgg tattgacact tgaggggcgc gactactgac agatgagggg
     5101 cgcgatcctt gacacttgag gggcagagtg ctgacagatg aggggcgcac ctattgacat
     5161 ttgaggggct gtccacaggc agaaaatcca gcatttgcaa gggtttccgc ccgtttttcg
     5221 gccaccgcta acctgtcttt taacctgctt ttaaaccaat atttataaac cttgttttta
     5281 accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa ggggggtgcc cccccttctc
     5341 gaaccctccc ggcccgctaa cgcgggcctc ccatcccccc aggggctgcg cccctcggcc
     5401 gcgaacggcc tcaccccaaa aatggcagcg ctggccaatt cgtgcgcgga acccctattt
     5461 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccttgataaa
     5521 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5581 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5641 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5701 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5761 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5821 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5881 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5941 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6001 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6061 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6121 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6181 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6241 ataaatctgg agccggtgag cgtggttctc gcggtatcat tgcagcactg gggccagatg
     6301 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6361 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6421 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6481 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6541 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6601 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6661 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6721 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6781 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6841 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6901 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6961 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7021 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7081 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7141 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7201 tggcagatcc tagatgtggc gcaacgatgc cggcgacaag caggagcgca ccgacttctt
     7261 ccgcatcaag tgttttggct ctcaggccga ggcccacggc aagtatttgg gcaaggggtc
     7321 gctggtattc gtgcagggca agattcggaa taccaagtac gagaaggacg gccagacggt
     7381 ctacgggacc gacttcattg ccgataaggt ggattatctg gacaccaagg caccaggcgg
     7441 gtcaaatcag gaataagggc acattgcccc ggcgtgagtc ggggcaatcc cgcaaggagg
     7501 gtgaatgaat cggacgtttg accggaaggc atacaggcaa gaactgatcg acgcggggtt
     7561 ttccgccgag gatgccgaaa ccatcgcaag ccgcaccgtc atgcgtgcgc cccgcgaaac
     7621 cttccagtcc gtcggctcga tggtccagca agctacggcc aagatcgagc gcgacagcgt
     7681 gcaactggct ccccctgccc tgcccgcgcc atcggccgcc gtggagcgtt cgcgtcgtct
     7741 tgaacaggag gcggcaggtt tggcgaagtc gatgaccatc gacacgcgag gaactatgac
     7801 gaccaagaag cgaaaaaccg ccggcgagga cctggcaaaa caggtcagcg aggccaagca
     7861 ggccgcgttg ctgaaacaca cgaagcagca gatcaaggaa atgcagcttt ccttgttcga
     7921 tattgcgccg tggccggaca cgatgcgagc gatgccaaac gacacggccc gctctgccct
     7981 gttcaccacg cgcaacaaga aaatcccgcg cgaggcgctg caaaacaagg tcattttcca
     8041 cgtcaacaag gacgtgaaga tcacctacac cggcgtcgag ctgcgggccg acgatgacga
     8101 actggtgtgg cagcaggtgt tggagtacgc gaagcgcacc cctatcggcg agccgatcac
     8161 cttcacgttc tacgagcttt gccaggacct gggctggtcg atcaatggcc ggtattacac
     8221 gaaggccgag gaatgcctgt cgcgcctaca ggcgacggcg atgggcttca cgtccgaccg
     8281 cgttgggcac ctggaatcgg tgtcgctgct gcaccgcttc cgcgtcctgg accgtggcaa
     8341 gaaaacgtcc cgttgccagg tcctgatcga cgaggaaatc gtcgtgctgt ttgctggcga
     8401 ccactacacg aaattcatat gggagaagta ccgcaagctg tcgccgacgg cccgacggat
     8461 gttcgactat ttcagctcgc accgggagcc gtacccgctc aagctggaaa ccttccgcct
     8521 catgtgcgga tcggattcca cccgcgtgaa gaagtggcgc gagcaggtcg gcgaagcctg
     8581 cgaagagttg cgaggcagcg gcctggtgga acacgcctgg gtcaatgatg acctggtgca
     8641 ttgcaaacgc tagggccttg tggggtcagt tccggctggg ggttcagcag ccagcgcctg
     8701 atctggggaa ccctgtggtt ggcatgcaca tacaaatgga cgaacggata aaccttttca
     8761 cgccctttta aatatccgat tattctaata aacgctcttt tctcttag
//
