LOCUS       sequence-415344-        8769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Premade nuclei membrane marker, pUBQ10:H2B:mTurquoise2:tHSP18.2M, 
            for braiding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8769)
  AUTHORS   Piepers M, Erbstein K, Reyes-Hernandez J, Song C, Tessi T, 
            Petrasiunaite V, Faerber N, Distel K, Maizel A
  TITLE     GreenGate 2.0: Backwards compatible addons for assembly of complex 
            transcriptional units and their stacking with GreenGate.
  JOURNAL   PLoS One. 2023 Sep 8;18(9):e0290097. doi: 
            10.1371/journal.pone.0290097. eCollection 2023.
  PUBMED    37682951
REFERENCE   2  (bases 1 to 8769)
  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"; journalName: "PLoS One.";
            date: "2023-09-8"; volume: "18(9):e0290097. doi"; pages: " 
            10.1371/journal.pone.0290097. eCollection 2023"
COMMENT     Sequence Label: pKD012
FEATURES             Location/Qualifiers
     source          1..8769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(326..1039,1040..1042,1043..1045))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(371..392)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     720..739
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     979..1000
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2434..2458
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3759..4388
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4817..5890
                     /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      5956..6150
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6386..6408)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6494..6634
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6549..6568
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6649..6666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6820..7408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6900..6919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7496..8311)
                     /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(7496..8311)
                     /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     8219..8238
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8744..8768
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacgaattg ctcgagcacc tgccgatata cgttctccta tggtcatgtg tcacttagtg
       61 aattccttat ctttaatcat attccatagt ccataccata gcacatacag tagttatatg
      121 ctgcagaaga gatccaacaa aacattcaca atggattata gaaacatttg tttattcatt
      181 ataatgagat cttacattca tttaatatta gaaaaagcca caaattcata acacaacaag
      241 ccaagaaaaa aacacaaact taagcacaca agctatttat ttgacacacc aaatatttca
      301 tcttcatctt catatgagct cgcagctact tgtacagctc gtccatgccg agagtgatcc
      361 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
      421 tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
      481 tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc
      541 ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata tagacgttgt
      601 cgctaaagta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
      661 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
      721 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
      781 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg aagcactgca
      841 cgccccagga cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
      901 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
      961 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     1021 acagctcctc gcccttgctc accatggatc cgcctccacc actagtgcca ccgctgccac
     1081 cgctagcacc tgaacttgaa gagccggaac cagatccgga ccctgaaccc gaacctgatc
     1141 ctgatccgga cccactacct gagccagatc cggagccaga acctgaagca gcagcggagc
     1201 ctgcagcggc cgccgatcca gagccgctgc cggaaccact ccctgagccc gatccagagc
     1261 cgcttccgga accactccct gatgcagacg cactagcggc tgtcgaacct gaagagctag
     1321 taaacttagt aacagcctta gttccttcag aaacagcatg cttagcaagt tcaccaggta
     1381 acactagtct aacagctgtc tgaatctccc tagaagtaat cgtaggtttc ttattatacc
     1441 tagctaattt tgaagcttct tgcgcaagtt tctcgaatat atcattgata aaactgttca
     1501 taatccccat agctttactt gagattccaa tatctggatg aacttgtttc aacaccttga
     1561 agatgtagat cttgtatgtc tctgtgcttt tcttgactcg tttcttcttc ttctcgactc
     1621 ctccggtgac tgcttctttt ggtagttttt ttccggcttt tggtttcttc tctgctggtg
     1681 ctttttcggc tactttctct tcttctgctg gagctttctc tgcgggcttt ttctctgctg
     1741 gtttctttcc ggcttttggt gccattgttt gttaatcaga aaaactcaga ttaatctaca
     1801 aattcgatcg cacaaactag aaactaacac cagatctaga tagaaatcac aaatcgaaga
     1861 gtaattattc gacaaaactc aaattatttg aacaaatcgg atgatatcta tgaaacccta
     1921 atcgagaatt aagatgatat ctaacgatca aacccagaaa atcgtcttcg atctaagatt
     1981 aacagaatct aaaccaaaga acatatacga aattgggatc gaacgaaaac aaaatcgaag
     2041 attttgagag aataaggaac acagaaattt accttgatca cggtagagag aattgagaga
     2101 aagtttttaa gattttgaga aattgaaatc tgaattgtga agaagaagag ctctttgggt
     2161 attgttttat agaagaagaa gaagaaaaga cgaggacgac taggtcacga gaaagctaag
     2221 gcggtgaagc aatagctaat aataaaatga cacgtgtatt gagcgttgtt tacacgcaaa
     2281 gttgtttttg gctaattgcc ttatttttag gttgaggaaa agtatttgtg ctttgagttg
     2341 ataaacacga ctcgtgtgtg ccggctgcaa ccactttgac gccgtttatt actgactcgt
     2401 cgaggttgac tgtagcaggt gccatggtaa ttctgacagg atatattggc gggtaaacct
     2461 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
     2521 cgttcgtcca tttgtatgtg catgccaacc acagggttcc cctcgggatc aaagtacttt
     2581 gatccaaccc ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcatc
     2641 tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc
     2701 ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg
     2761 gagacattac gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag
     2821 caccgacgac caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa
     2881 gctgttttcc gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct
     2941 tgaccaccta cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag
     3001 cacccgcgac ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag
     3061 cctggcagag ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt
     3121 cgccggcatt gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga
     3181 ggccgccaag gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat
     3241 cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact
     3301 gcttggcgtg catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc
     3361 caccgaggcc aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct
     3421 ggcggccgcc gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag
     3481 gacgaaccgt ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg
     3541 ttcgagccgc ccgcgcacct ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct
     3601 gatgccaagc tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt
     3661 ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg
     3721 atgcgatgag taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta
     3781 accagaaagg cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac
     3841 tcgccggggc cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg
     3901 cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc
     3961 gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg
     4021 cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa
     4081 gcccttacga catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg
     4141 tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca
     4201 tcggcggtga ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta
     4261 tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag
     4321 aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac
     4381 tcatttgagt taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg
     4441 ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc
     4501 catgaagcgg gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc
     4561 ggtacgccaa ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga
     4621 gtaaatgagc aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg
     4681 aaaatcaaga acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg
     4741 gttggccagg cgtaagcggc tgggttgcct gccggccctg caatggcact ggaaccccca
     4801 agcccgagga atcggcgtga gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc
     4861 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat
     4921 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga
     4981 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga
     5041 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat
     5101 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg
     5161 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt
     5221 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata
     5281 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact
     5341 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat
     5401 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct
     5461 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac
     5521 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga
     5581 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat
     5641 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt
     5701 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac
     5761 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg
     5821 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc
     5881 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg
     5941 aaaaggactc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa
     6001 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt
     6061 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa
     6121 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct
     6181 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc
     6241 tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg
     6301 cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg
     6361 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggtg acggtcacag
     6421 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6481 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     6541 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6601 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     6661 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6721 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6781 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6841 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6901 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6961 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7021 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7081 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7141 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7201 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7261 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7321 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7381 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7441 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgcattcta ggtgattaga
     7501 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     7561 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     7621 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     7681 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     7741 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     7801 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     7861 gagcgagtcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     7921 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     7981 ctaatacctg gaatgctgtt ttccctggga tcgcagtggt gagtaaccat gcatcatcag
     8041 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     8101 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     8161 ctggcgcatc gggcttccca tacaatcggt agattgtcgc acctgattgc ccgacattat
     8221 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggccttg
     8281 agcaagacgt ttcccgttga atatggctca taacagaact tattatttcc ttcctctttt
     8341 ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc caattcactg
     8401 ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt gttttcaaag
     8461 ttggcgtata acatagtatc gacggagccg attttgaaac cgcggtgatc acaggcagca
     8521 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     8581 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
     8641 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
     8701 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     8761 gtgtaaaca
//
