LOCUS       sequence-386926-        5862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YFP expression cassette: CaMV35S::TMV::YFP::35sT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5862)
  AUTHORS   Cai YM, Witham S, Patron NJ
  TITLE     Tuning Plant Promoters Using a Simple Split Luciferase Method to 
            Assess Transcription Factor-DNA Interactions.
  JOURNAL   ACS Synth Biol. 2023 Nov 17;12(11):3482-3486. doi: 
            10.1021/acssynbio.3c00094. Epub 2023 Oct 19.
  PUBMED    37856867
REFERENCE   2  (bases 1 to 5862)
  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"; doi: 
            "10.1021/acssynbio.3c00094"; journalName: "ACS Synth Biol"; date: 
            "2023-11-17- 17"; volume: "12"; issue: "11"; pages: "3482-3486"
COMMENT     Sequence Label: pEPOR1CB0068
FEATURES             Location/Qualifiers
     source          1..5862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(271..1062,1063..1131))
                     /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     894..913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1132..1236)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1323..1954
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    2046..2070
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2256..2601
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2544..2564
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             join(2692..2694,2695..2697,2698..3408)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2737..2758)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2998..3017)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3345..3366
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3491..3665
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3710..3734
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3890..5038)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(join(5374..5862,1..100))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5454..5473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
       61 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
      301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
      361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg
      421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
      481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
      541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1141 ttttcaatat tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga
     1201 atgtatttag aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt
     1261 tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg
     1321 ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc
     1381 gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta
     1441 aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct
     1501 gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc
     1561 ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata
     1621 ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca
     1681 ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg
     1741 cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc
     1801 tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc ggccgcggtg
     1861 tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag
     1921 cccagcggcg agggcaacca gcccggtgag cgtcgcaaag gagatcctga tctgactgat
     1981 gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg
     2041 gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca
     2101 ttgcggacgt ttttaatgta ctggggtgga tgcagtgggc cccactctgt gaagacaatt
     2161 acgaattccc atggggaggt caacatggtg gagcacgaca ctctggtcta ctccaaaaat
     2221 gtcaaagata cagtctcaga agatcaaagg gctattgaga cttttcaaca aaggataatt
     2281 tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta
     2341 gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa
     2401 gatctctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa
     2461 aaagaagagg ttccaaccac gtctacaaag caagtggatt gatgtgacat ctccactgac
     2521 gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt
     2581 tcatttcatt tggagaggac acgctcgagt ataagagctc atttttacaa caattaccaa
     2641 caacaacaaa caacaaacaa cattacaatt acatttacaa ttatcgatac aatggtgagc
     2701 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2761 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     2821 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2881 accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     2941 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3001 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3061 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3121 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3181 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3241 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3301 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3361 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagaa gaagagaaag
     3421 gtaaagaaga gaaaggtaaa gaagagaaag gtatgagctt ctctagctag agtcgatcga
     3481 caagctcgag tttctccata ataatgtgtg agtagttccc agataaggga attagggttc
     3541 ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg
     3601 taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc
     3661 agatcgctca gattgtcttc tgcacgaagt ggtttaaact atcagtgttt gacaggatat
     3721 attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg
     3781 cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttccccag
     3841 atcaggcgct ggctgctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc
     3901 aatgcaccag gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt
     3961 cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca
     4021 tgaggcggaa ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga
     4081 acatccgtcg ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt
     4141 ggtcgccagc aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt
     4201 tcttgccacg gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa
     4261 cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct
     4321 tcgtgtaata ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga
     4381 aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc caacacctgc tgccacacca
     4441 gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga
     4501 cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga
     4561 acagggcaga gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa
     4621 tatcgaacaa ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg
     4681 cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg
     4741 tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt
     4801 caagacgacg cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt
     4861 gcacgctgtc gcgctcgatc ttggccgtag cttgctggac catcgagccg acggactgga
     4921 aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg
     4981 aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc
     5041 accctccttg cgggattgcc ccgactcacg ccggggcaat gtgcccttat tcctgatttg
     5101 acccgcctgg tgccttggtg tccagataat ccaccttatc ggcaatgaag tcggtcccgt
     5161 agaccgtctg gccgtccttc tcgtacttgg tattccgaat cttgccctgc acgaatacca
     5221 gcgacccctt gcccaaatac ttgccgtggg cctcggcctg agagccaaaa cacttgatgc
     5281 ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt gcgccacatc taggatctgc
     5341 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5401 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5461 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5521 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5581 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5641 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5701 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5761 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5821 atttggtatc tgcgctctgc tgaagccagt taccttcgga aa
//
