LOCUS       sequence-299499-        8600 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 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 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: pEPYC1CB0305
FEATURES             Location/Qualifiers
     source          1..8600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(1044..1112,1113..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             join(7159..7161,7162..7164,7165..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
//
