LOCUS       sequence-386944-        7050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1 luciferase reporter: MinSyn_301 reporter 
            [MinSyn_301::TMV::LucF::flag::ocsT].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7050)
  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 7050)
  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: pEPYC1CB0597
FEATURES             Location/Qualifiers
     source          1..7050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    42..66
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(222..1370)
                     /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(1706..2294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1786..1805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2465..3256,3257..3325))
                     /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     3088..3107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3326..3430)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3517..4148
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4240..4264
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    4532..4587
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     CDS             4589..6238
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(4655..4673)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             6302..6325
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      6336..7043
                     /label=OCS terminator
                     /note="octopine synthase terminator"
ORIGIN
        1 gcaattgtct tctgcacgaa gtggtttaaa ctatcagtgt ttgacaggat atattggcgg
       61 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa
      121 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc agatcaggcg
      181 ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc
      241 aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct
      301 tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg
      361 aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt
      421 cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca
      481 gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca
      541 cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg
      601 gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa
      661 taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc
      721 ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca
      781 tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa
      841 atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca
      901 gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac
      961 aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg
     1021 gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata
     1081 gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcaagacga
     1141 cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg
     1201 tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg gaaggtttcg
     1261 cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc
     1321 gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct
     1381 tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt tgacccgcct
     1441 ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc gtagaccgtc
     1501 tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac cagcgacccc
     1561 ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat gcggaagaag
     1621 tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggatct gccaggaacc
     1681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2581 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa
     2641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3361 aaggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3421 gggttccgcg cacgaattgg ccagcgctgc catttttggg gtgaggccgt tcgcggccga
     3481 ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag
     3541 ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa
     3601 caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa
     3661 acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca
     3721 ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc
     3781 atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg
     3841 tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg
     3901 gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg
     3961 gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt
     4021 ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc acacggcttc gacggcgttt
     4081 ctggcgcgtt tgcagggcca tagacggccg ccagcccagc ggcgagggca accagcccgg
     4141 tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg cctgtatcga gtggtgattt
     4201 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt
     4261 aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg
     4321 tggatgcagt gggccccact ctgtgaagac aatgccgaat tcggatccgg agggaggcgt
     4381 gtcgttttag tgaggcttac gtctgggatt acgtctactt acgtctggga ttacgtctac
     4441 ttacgtctgg gattacgtct actatataag gttttgctat tcattgaaag cagtagtgac
     4501 tgatttgtat ataatgtact gtatttttac aacaattacc aacaacaaca aacaacaaac
     4561 aacattacaa ttactattta caattacaat ggaggacgcc aaaaacataa agaaaggccc
     4621 ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa
     4681 gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat
     4741 cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg
     4801 gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc
     4861 ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga
     4921 acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa
     4981 ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat
     5041 catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca
     5101 tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac
     5161 aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc
     5221 tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat
     5281 cattccggat actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac
     5341 tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga
     5401 actgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg tgccaaccct
     5461 attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga
     5521 aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt
     5581 ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat
     5641 tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc
     5701 gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg
     5761 tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt
     5821 gattgacaag gatggatggc tacattctgg agacatagct tactgggacg aggacgaaca
     5881 cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc
     5941 cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg
     6001 tctgcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa
     6061 gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa
     6121 gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga
     6181 cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtggg
     6241 ttcggattat aaggaccatg acggagacta taaggaccat gacctcgacg ctgcagcagc
     6301 ggattataag gacgatgacg ataagtgagc ttgtcctgct ttaatgagat atgcgagaag
     6361 cctatgatcg catgatattt gctttcaatt ctgttgtgca cgttgtaaaa aacctgagca
     6421 tgtgtagctc agatccttac cgccggtttc ggttcattct aatgaatata tcacccgtta
     6481 ctatcgtatt tttatgaata atattctccg ttcaatttac tgattgtacc ctactactta
     6541 tatgtacaat attaaaatga aaacaatata ttgtgctgaa taggtttata gcgacatcta
     6601 tgatagagcg ccacaataac aaacaattgc gttttattat tacaaatcca attttaaaaa
     6661 aagcggcaga accggtcaaa cctaaaagac tgattacata aatcttattc aaatttcaaa
     6721 agtgccccag gggctagtat ctacgacaca ccgagcggcg aactaataac gctcactgaa
     6781 gggaactccg gttccccgcc ggcgcgcatg ggtgagattc cttgaagttg agtattggcc
     6841 gtccgctcta ccgaaagtta cgggcaccat tcaacccggt ccagcacggc ggccgggtaa
     6901 ccgacttgct gccccgagaa ttatgcagca tttttttggt gtatgtgggc cccaaatgaa
     6961 gtgcaggtca aaccttgaca gtgacgacaa atcgttgggc gggtccaggg cgaattttgc
     7021 gacaacatgt cgaggctcag caggaccgct
//
