LOCUS       sequence-387014-        8400 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 8400)
  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 8400)
  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: pEPSW1KN0024
FEATURES             Location/Qualifiers
     source          1..8400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      315..509
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(745..767)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    853..993
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     908..927
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1008..1025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1179..1767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1259..1278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1855..2670)
                     /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(1855..2670)
                     /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     2578..2597
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3103..3127
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             4305..4817
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             4878..4901
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      4917..5169
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5193..5217
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6518..7147
                     /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(7576..8400,1..249)
                     /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 ggccgttttc tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg
       61 ttcaagacga tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc
      121 gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg
      181 caggctggcc cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc
      241 ggttcctaat gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga
      301 aaaggactct ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac
      361 cggaacccgt acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg
      421 actgatataa aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa
      481 actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaagtgctg
      541 caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct
      601 atcgcggccg ctggccgctc aaaaatggct ggcctacggc caggcaatct accagggcgc
      661 ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac atcaaggcac cctgcctcgc
      721 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggtga cggtcacagc
      781 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      841 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
      901 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
      961 cacagatgcg taaggagaaa ataccgcatc aggcgctcat ccgcttcctc gctcactgac
     1021 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1081 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1141 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1201 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1261 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1321 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1381 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1441 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1501 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1561 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     1621 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1681 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1741 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1801 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcattctag gtgattagaa
     1861 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     1921 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     1981 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     2041 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     2101 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     2161 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     2221 agcgagtcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     2281 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     2341 taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg
     2401 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     2461 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     2521 tggcgcatcg ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc
     2581 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga
     2641 gcaagacgtt tcccgttgaa tatggctcat aacagaactt attatttcct tcctcttttc
     2701 tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt
     2761 tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt
     2821 tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa
     2881 cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc
     2941 cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct
     3001 tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga
     3061 ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg
     3121 tgtaaacata acgaattgct cttcaatgag gagtactact gcaaattttt aaaaagttaa
     3181 tattcgactt tataaaagag atgagagtca acaaaacaaa aagcaactaa ttttggacgt
     3241 acgaaatgct ttgtaaatac attgatatca cttatgttta taatatcact catcccaagt
     3301 gttggtcaac atttctttga taaaaattaa actattcaat agaacgaaaa ccacaaaaaa
     3361 attataaaga attattgata acaactcttc atgttaatct tatatatttt tttaaaaata
     3421 tttgttttta ggttttaata caaacaacat gattctagtt tattaaacta attaacgcaa
     3481 cttgatggat tataaaaata tagcttaact aatccatcaa attaacaagt gaatcgcact
     3541 ataattctaa aagaatcagg taggtccagg atttgtcttt tctccacttc tcaatttccc
     3601 attaataatc ccgaaaaggc attacgtaat aattaatatt ttacgctgta gtattatttt
     3661 atgataaaca tttccaaatt ttccaacact taatcataat ttaaggatta aaggaaattt
     3721 gattggtgca ttcccaccac gagtttgctt aacgtgtcgt taaatgcagc cgtcggactt
     3781 gacaatgcaa aaatgctaca taatctaaat aattaactct cgcaataaga ttcactaata
     3841 ataacatttt taacgacgat atataatttt acaataagaa aaaaaatatg aatctaatat
     3901 ttatttaata ttaatttact ccattatctg aataatctag aatatgtcaa atatcctttt
     3961 attattttgt agctgtcgaa actatggtga ataaaatctc gaggaatata ttagaaaaca
     4021 aaaaagatct agaggaataa taatatttta aatatttggg gcacagttgt attgtgaagc
     4081 agagaatcgg ttctcaatat attatccgaa aatgtcttaa ttttaaattt tgttttttgt
     4141 ttgattctgc gatattggaa tcacacactc tgccaatttg acctttctct ctcgcttctc
     4201 tttctctatc caatcggaaa gagatcaacc aacgatagag gagaagaaga acttgcatac
     4261 gcaaaaaaac tttcccggga aaattccaga aactgctttg gaaaatggta ttcacactcg
     4321 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     4381 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     4441 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     4501 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     4561 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     4621 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     4681 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     4741 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     4801 gcgaacgcat tctggcttcg gattataagg accatgacgg agactataag gaccatgacc
     4861 tcgacgctgc agcagcggat tataaggacg atgacgataa gtgagcttgt caagcagatc
     4921 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
     4981 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
     5041 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
     5101 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
     5161 tactagatcg acgctagcat gaagagcaat tctgacagga tatattggcg ggtaaaccta
     5221 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
     5281 gttcgtccat ttgtatgtgc atgccaacca cagggttccc ctcgggatca aagtactttg
     5341 atccaacccc tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcatct
     5401 gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc
     5461 tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg
     5521 agacattacg ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc
     5581 accgacgacc aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag
     5641 ctgttttccg agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt
     5701 gaccacctac gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc
     5761 acccgcgacc tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc
     5821 ctggcagagc cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc
     5881 gccggcattg ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag
     5941 gccgccaagg cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc
     6001 gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg
     6061 cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc
     6121 accgaggcca ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg
     6181 gcggccgccg agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg
     6241 acgaaccgtt tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt
     6301 tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg
     6361 atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc
     6421 taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga
     6481 tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa
     6541 ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact
     6601 cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc
     6661 ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg
     6721 cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc
     6781 ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag
     6841 cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt
     6901 cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat
     6961 cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat
     7021 cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga
     7081 acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact
     7141 catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc
     7201 cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc
     7261 atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg
     7321 gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag
     7381 taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga
     7441 aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg
     7501 ttggccaggc gtaagcggct gggttgcctg ccggccctgc aatggcactg gaacccccaa
     7561 gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg
     7621 ctgggtgatg acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc
     7681 gaggcagaag cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa
     7741 tcccggcaac cgccggcagc cggtgcgccg tcgattagga agccgcccaa gggcgacgag
     7801 caaccagatt ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc
     7861 atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc
     7921 tacgagcttc cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg
     7981 tgggattacg acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac
     8041 cgggaaggga agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc
     8101 aagttctgcc ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt
     8161 cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg
     8221 gtgacggtat ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc
     8281 gggcggccgg agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa
     8341 ggcaagaacc cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc
//
