LOCUS       sequence-387012-        8262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1 luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8262)
  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 8262)
  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: pEPSW1KN0025
FEATURES             Location/Qualifiers
     source          1..8262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      314..508
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(744..766)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    852..992
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     907..926
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1007..1024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1178..1766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1258..1277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1854..2669)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1854..2669)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2577..2596
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3102..3126
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             4166..4678
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             4739..4762
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      4778..5030
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5054..5078
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6379..7008
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(7437..8262,1..248)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt
       61 tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg
      121 tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc
      181 aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg
      241 gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa
      301 aaggactctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc
      361 ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga
      421 ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa
      481 ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagtgctgc
      541 aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta
      601 tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg
      661 gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg
      721 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggtgac ggtcacagct
      781 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      841 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
      901 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
      961 acagatgcgt aaggagaaaa taccgcatca ggcgctcatc cgcttcctcg ctcactgact
     1021 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1081 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     1621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcattctagg tgattagaaa
     1861 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1921 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1981 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2041 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2101 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2161 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2221 gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2281 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2341 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     2401 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2461 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2521 ggcgcatcgg gcttcccata caatcggtag attgtcgcac ctgattgccc gacattatcg
     2581 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag
     2641 caagacgttt cccgttgaat atggctcata acagaactta ttatttcctt cctcttttct
     2701 acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt
     2761 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt
     2821 ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac
     2881 gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc
     2941 ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt
     3001 acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat
     3061 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt
     3121 gtaaacataa cgaattgctc ttcaatgagg agcctataaa atccctaatg agcacatcaa
     3181 ccttgaatct tctctaataa accttttttc caaacacgca ctaaaccaaa aattaacatc
     3241 tcaagaggaa accatttaaa aaaaaaacag agttagatta agatcaacaa atatagttga
     3301 aaagaacata tgttaagcaa cattaccata aattcataat tagtagtgat taaaacttaa
     3361 gaatactaaa gtatgaatat gaaaattgtt gtattttttt gtagtatgta atatgcggaa
     3421 acttggatgt tatcctattc attgaaattc atatatgtat gtagttatca ttttttaaat
     3481 atattaaaaa attaaaaagt aataatacta taaacattca aattatgaaa taaatatctc
     3541 atactaatca ctgctaattt tttttccaat gtttctcttt agtatttctc tttagtatgg
     3601 gagtgacaaa cttaatatga aatctatctc cctacatgca aaaatcgaat ctcttttatt
     3661 attaatgaag tgtatggtcg gtgtcaataa ctaatatgtc atatttttta cagactgttc
     3721 agatatgata aaaatagttt ttttccaacc aaatggttgg tcttctgagt gtatttgaaa
     3781 acgacattga agaattaata tatttttttt taattttagt tttttatagt acaaatatta
     3841 aaacaaacaa tcctaccata tcataacatt tgtaaataac attttaagtt ttgttttgag
     3901 ttttaattaa ttttctatga caaaaaaatg aagtcaatag actaagtgaa tcatatagta
     3961 taaataaaca caatttaaat agtttcaaat aaatttagaa agaataaaac aaatagaaat
     4021 cagaaggtgt ctgtttcctc ctcgcaacat acgatcaaag agaaacaact tgacccttta
     4081 cattgctcaa gagctcatct cttccctcta caaaaatggc cgcacgtctc caaccttctc
     4141 ccaactcctt cttccgccat catcaatggt attcacactc gaagatttcg ttggggactg
     4201 gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg tgtccagttt
     4261 gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa
     4321 tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat
     4381 gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc actttaaggt
     4441 gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga tcgactattt
     4501 cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg taacagggac
     4561 cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg gctccctgct
     4621 gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca ttctggcttc
     4681 ggattataag gaccatgacg gagactataa ggaccatgac ctcgacgctg cagcagcgga
     4741 ttataaggac gatgacgata agtgagcttg tcaagcagat cgttcaaaca tttggcaata
     4801 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     4861 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     4921 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     4981 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gacgctagca
     5041 tgaagagcaa ttctgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt
     5101 agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg
     5161 catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct
     5221 atagtgcagt cggcttctga cgttcagtgc agccgtcatc tgaaaacgac atgtcgcaca
     5281 agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt
     5341 gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca
     5401 agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga
     5461 ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca
     5521 ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg
     5581 acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca
     5641 ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg
     5701 acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg
     5761 agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg
     5821 tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga
     5881 tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga
     5941 ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg
     6001 gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac
     6061 gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac
     6121 cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacct
     6181 ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg
     6241 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt
     6301 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca
     6361 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc
     6421 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg
     6481 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa
     6541 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc
     6601 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg
     6661 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc
     6721 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa
     6781 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag
     6841 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac
     6901 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc
     6961 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta
     7021 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca
     7081 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc
     7141 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca
     7201 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa
     7261 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc
     7321 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc
     7381 tgggttgcct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga
     7441 gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg
     7501 agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg
     7561 gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag
     7621 ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc
     7681 cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc
     7741 gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc
     7801 acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac
     7861 tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca
     7921 agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg
     7981 atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg
     8041 ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg
     8101 aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg
     8161 agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc
     8221 tgacggttca ccccgattac tttttgatcg atcccggcat cg
//
