LOCUS       sequence-249656-        8962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Napin Promoter driven dsRED expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8962)
  AUTHORS   Wu G, Rossidivito G, Hu T, Berlyand Y, Poethig RS
  TITLE     Traffic lines: new tools for genetic analysis in Arabidopsis 
            thaliana.
  JOURNAL   Genetics. 2015 May;200(1):35-45. doi: 10.1534/genetics.114.173435. 
            Epub 2015 Feb 23.
  PUBMED    25711279
REFERENCE   2  (bases 1 to 8962)
  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: 
            "10.1534/genetics.114.173435"; journalName: "Genetics"; date: 
            "2015-05"; volume: "200"; issue: "1"; pages: "35-45"
COMMENT     Sequence Label: pNapin-dsRED
FEATURES             Location/Qualifiers
     source          1..8962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1219..1848
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2277..3350
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3416..3610
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3846..3868)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3954..4094
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4009..4028
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4109..4126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4280..4868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4360..4379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4955..5749)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6174..6198
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    6276..6450
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     terminator      8345..8597
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(8637..8654)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8637..8653)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8646..8668)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    8856..8880
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct
       61 atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca
      121 agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt
      181 gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca
      241 agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga
      301 ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca
      361 ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg
      421 acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca
      481 ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg
      541 acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg
      601 agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg
      661 tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga
      721 tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga
      781 ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg
      841 gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac
      901 gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac
      961 cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt
     1021 ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg
     1081 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt
     1141 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca
     1201 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc
     1261 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg
     1321 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa
     1381 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc
     1441 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg
     1501 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc
     1561 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa
     1621 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag
     1681 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac
     1741 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc
     1801 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta
     1861 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca
     1921 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc
     1981 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca
     2041 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa
     2101 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc
     2161 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc
     2221 tgggttgcct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga
     2281 gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg
     2341 agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg
     2401 gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag
     2461 ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc
     2521 cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc
     2581 gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc
     2641 acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac
     2701 tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca
     2761 agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg
     2821 atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg
     2881 ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg
     2941 aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg
     3001 agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc
     3061 tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc
     3121 tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac
     3181 gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt
     3241 caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag
     3301 tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc
     3361 agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggtctc tttcctgtgg
     3421 atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga
     3481 acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa
     3541 aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg
     3601 cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc
     3661 ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct
     3721 caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc
     3781 cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt
     3841 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3901 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc
     3961 atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc
     4021 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4081 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4141 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4201 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4261 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4321 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4381 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4441 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4501 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4561 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4621 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4681 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4741 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4801 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4861 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgc tgatgaatcc
     4921 cacgttaagg gattttggtc atgcattcta ggtactaaaa caattcatcc agtaaaatat
     4981 aatattttat tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat agctcgacat
     5041 actgttcttc cccgatatcc tccctgatcg accggacgca gaaggcaatg tcataccact
     5101 tgtccgccct gccgcttctc ccaagatcaa taaagccact tactttgcca tctttcacaa
     5161 agatgttgct gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg
     5221 tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg agtgtcttct tcccagtttt
     5281 cgcaatccac atcggccaga tcgttattca gtaagtaatc caattcggct aagcggctgt
     5341 ctaagctatt cgtataggga caatccgata tgtcgatgga gtgaaagagc ctgatgcact
     5401 ccgcatacag ctcgataatc ttttcagggc tttgttcatc ttcatactct tccgagcaaa
     5461 ggacgccatc ggcctcactc atgagcagat tgctccagcc atcatgccgt tcaaagtgca
     5521 ggacctttgg aacaggcagc tttccttcca gccatagcat catgtccttt tcccgttcca
     5581 catcataggt ggtcccttta taccggctgt ccgtcatttt taaatatagg ttttcatttt
     5641 ctcccaccag cttatatacc ttagcaggag acattccttc cgtatctttt acgcagcggt
     5701 atttttcgat cagttttttc aattccggtg atattctcat tttagccatt tattatttcc
     5761 ttcctctttt ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc
     5821 caattcactg ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt
     5881 gttttcaaag ttggcgtata acatagtatc gacggagccg attttgaaac cgcggtgatc
     5941 acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg
     6001 tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa
     6061 gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta
     6121 tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg
     6181 atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt
     6241 ttaatgtact gaattaacgc cgaattaatt cgggggatct ggattttagt actggatttt
     6301 ggttttagga attagaaatt ttattgatag aagtatttta caaatacaaa tacatactaa
     6361 gggtttctta tatgctcaac acatgagcga aaccctatag gaaccctaat tcccttatct
     6421 gggaactact cacacattat tatggagaaa ctcgaggatc tagtaacata cctgcaggtc
     6481 gactctagag gatccccggg taccgagctc gaattcgatt cggagctctc ttcatcggtg
     6541 attgattcct ttaaagactt atgtttctta tcttgcttct gaggcaagta ttcagttacc
     6601 agttaccact tatattctgg actttctgac tgcatcctca tttttccaac attttaaatt
     6661 tcactattgg ctgaatgctt cttctttgag gaagaaacaa ttcagatggc agaaatgtat
     6721 caaccaatgc atatatacaa atgtacctct tgttctcaaa acatctatcg gatggttcca
     6781 tttgctttgt catccaatta gtgactactt tatattattc actcctcttt attactattt
     6841 tcatgcgagg ttgccatgta cattatattt gtaaggattg acgctattga gcgtttttct
     6901 tcaattttct ttattttaga catgggtatg aaatgtgtgt tagagttggg ttgaatgaga
     6961 tatacgttca agtgaagtgg cataccgttg tcgagtaagg atgacctacc cattcttgag
     7021 acaaatgtta cattttagta tcagagtaaa atgtgtacct ataactcaaa ttcgattgac
     7081 atgtatccat tcaacataaa attaaaccag cctgcacctg catccacatt tcaagtattt
     7141 tcaaaccgtt cggctcctat ccaccgggtg taacaagacg gattccgaat ttggaagatt
     7201 ttgactcaaa ttcccaattt atattgaccg tgactaaatc aactttaact tctataattc
     7261 tgattaagct cccaatttat attcccaacg gcactacctc caaaatttat agactctcat
     7321 ccccttttaa accaacttag taaacgctct tttttttaat tttatgaagt taagttttta
     7381 ccttgttttt aaaaagaatc gttcataaga tgccatgcca gaacattagc tacacgttac
     7441 acatagcatg cagccgcgga gaattgtttt tcttcgccac ttgtcactcc cttcaaacac
     7501 ctaagagctt ctctctcaca gcacacacat acaatcacat gcgtgcatgc attattacac
     7561 gtgatcgcca tgcaaatctc ctttatagtc tataaattaa ctcatccgct tcactcttta
     7621 ctcaaaccaa aactcatcaa tacaaacaag attaaaaaca taccccatgc ggtcttccaa
     7681 gaatgttatc aaggagttca tgaggtttaa ggttcgcatg gaaggaacgg tcaatgggca
     7741 cgagtttgaa atagaaggcg aaggagaggg gaggccatac gaaggccaca ataccgtaaa
     7801 gcttaaggta accaaggggg gacctttgcc atttgcttgg gatattttgt caccacaatt
     7861 tcagtatgga agcaaggtat atgtcaagca ccctgccgac ataccagact ataaaaagct
     7921 gtcatttcct gaaggattta aatgggaaag ggtcatgaac tttgaagacg gtggcgtcgt
     7981 tactgtaacc caggattcca gtttgcagga tggctgtttc atctacaagg tcaagttcat
     8041 tggcgtgaac tttccttccg atggacctgt tatgcaaaag aagacaatgg gctgggaagc
     8101 cagcactgag cgtttgtatc ctcgtgatgg cgtgttgaaa ggagagattc ataaggctct
     8161 gaagctgaaa aacggtggtc attacctagt tgaattcaaa agtatttaca tggcaaagaa
     8221 gcctgtgcag ctaccagggt actactatgt tgactccaaa ctggatataa caagccacaa
     8281 cgaagactat acaatcgttg agcagtatga aagaaccgag ggacgccacc atctgttcct
     8341 ttaagatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
     8401 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
     8461 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
     8521 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
     8581 atctatgtta ctagatcctc gcctctagag tcgacctgca ggcatgcaag cttggcactg
     8641 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     8701 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     8761 tcccaacagt tgcgcagcct gaatggcgaa tgctagagca gcttgagctt ggatcagatt
     8821 gtcgtttccc gccttcagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
     8881 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
     8941 tccgttcgtc catttgtatg tg
//
