LOCUS       sequence-318556-        8663 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 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 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: pEPYC1CB0478
FEATURES             Location/Qualifiers
     source          1..8663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(5148..5150,5151..5153,5154..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             join(7633..7701,7702..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
//
