LOCUS       sequence-387015-        8369 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 8369)
  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 8369)
  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: pEPSW1KN0022
FEATURES             Location/Qualifiers
     source          1..8369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1113..1307
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(1543..1565)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1651..1791
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1706..1725
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1806..1823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1977..2565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2057..2076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2653..3468)
                     /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(2653..3468)
                     /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     3376..3395
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3901..3925
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5072..5584
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             5645..5668
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      5684..5936
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5960..5984
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7285..7914
                     /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(8343..8369,1..1047)
                     /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 gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc
       61 gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca agcggccgct
      121 gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag
      181 ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga cgtgggcacc
      241 cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga
      301 gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc cgcagggccg
      361 gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc ccatctaacc
      421 gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt gttccgtcca
      481 cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca gaaagacgac
      541 ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg tacgaagaag
      601 gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag ccgctacaag
      661 atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc tgattggatg
      721 taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc cgattacttt
      781 ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag
      841 gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc cggagagttc
      901 aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat
      961 ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc
     1021 gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta
     1081 gcaggggaaa aaggtcgaaa aggactcttt cctgtggata gcacgtacat tgggaaccca
     1141 aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac
     1201 cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac
     1261 tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag
     1321 cgcacagccg aagtgctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc
     1381 gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca
     1441 ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat
     1501 caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     1561 cccggtgacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     1621 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     1681 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     1741 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcatcc
     1801 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1861 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1921 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2161 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2341 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2401 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2461 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2521 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2581 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2641 cattctaggt gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     2701 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     2761 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     2821 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     2881 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     2941 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     3001 cattcgtgat tgcgcctgag cgagtcgaaa tacgcgatcg ctgttaaaag gacaattaca
     3061 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     3121 tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag
     3181 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     3241 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     3301 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc
     3361 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     3421 atttaatcgc ggccttgagc aagacgtttc ccgttgaata tggctcataa cagaacttat
     3481 tatttccttc ctcttttcta cagtatttaa agatacccca agaagctaat tataacaaga
     3541 cgaactccaa ttcactgttc cttgcattct aaaaccttaa ataccagaaa acagcttttt
     3601 caaagttgtt ttcaaagttg gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc
     3661 ggtgatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga
     3721 tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat
     3781 gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct
     3841 gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg
     3901 tggcaggata tattgtggtg taaacataac gaattgctct tcaatgagga gcaataatta
     3961 aaccataaat ggggccactt tcttggaaag ttcgtattac gagttaacgc gttaaaacgc
     4021 agctgtaatt cgtaatctcc tgatagtgat gataggtacg tatatagtca atatataaaa
     4081 gaaaattgat atctcacaac ctaaaaaaat actataggat attttagcat cactcctcga
     4141 tttgctcatc aactggcgtc cgtgtgtata atatatatac tctcctggtt aattaattcc
     4201 tagcaaaatt gactccaatc ggtaatttat tttagtttga tagtttagtt cactcggttt
     4261 atattttgga caaattctag aaagaatcta acacgttaat aaacttggag agaataaatc
     4321 taattgtaaa atggaccctt gtgaaattaa tgttttttgt catttaaatt gtagaataaa
     4381 aaaatataca tttttctttg taacaagaac atgcaattga taaaaattta cgtgatcgca
     4441 taattaattg ttgttttgtg gttcctcaat aaaaaaagcc aatcaaaatc attatacgtt
     4501 tatgtatttt tcctctaaat accaagaata ttagttatac atgatttatt ttttataaga
     4561 aaatgcaaaa atagcaaaag gttaagatga ggccatgttc aggttcattg ttcgaacttc
     4621 gaacaattgg gcaaagcctt aacatgcatc catattcgaa taaataacta agctccaaag
     4681 atttatgttc actagtttct ttttttttct ttttttaatt tgaatggaac aaaaatgtat
     4741 taatgccaat tgctaggata aaggaaaaat tagaagaaaa acacctgtca tgcaccacaa
     4801 tgtcacattc atacacaaga aaccatacta tatttcattt gaccctattt tagatttata
     4861 tcaataaata atatttcatc tcttcctctc ccatacccct cccaaatcaa tccatctcta
     4921 tatataatct tctcagctca cactaatcct catcaaaaaa tctctatcca tccacaacaa
     4981 caacaaaaac tcaagaacaa tatctccact tgatcactac cacattgtcc aatcactcta
     5041 caaagcctgt acgtacacaa caacattacc aatggtattc acactcgaag atttcgttgg
     5101 ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc
     5161 cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg tcctgagcgg
     5221 tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcggcga
     5281 ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg atcatcactt
     5341 taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga acatgatcga
     5401 ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac
     5461 agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc ccgacggctc
     5521 cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg aacgcattct
     5581 ggcttcggat tataaggacc atgacggaga ctataaggac catgacctcg acgctgcagc
     5641 agcggattat aaggacgatg acgataagtg agcttgtcaa gcagatcgtt caaacatttg
     5701 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     5761 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     5821 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     5881 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcgacg
     5941 ctagcatgaa gagcaattct gacaggatat attggcgggt aaacctaaga gaaaagagcg
     6001 tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg
     6061 tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc caacccctcc
     6121 gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcatctgaa aacgacatgt
     6181 cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg
     6241 tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca
     6301 tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg
     6361 acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga
     6421 agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc
     6481 ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac
     6541 tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt
     6601 gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg
     6661 agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc
     6721 gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg
     6781 agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc
     6841 gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc
     6901 ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga
     6961 atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt
     7021 cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc
     7081 gcacctctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc
     7141 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg
     7201 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa
     7261 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg
     7321 tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat
     7381 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa
     7441 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc
     7501 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg
     7561 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata
     7621 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
     7681 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
     7741 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
     7801 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
     7861 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
     7921 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
     7981 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
     8041 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
     8101 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
     8161 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
     8221 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
     8281 agcggctggg ttgcctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
     8341 gcgtgagcgg tcgcaaacca tccggcccg
//
