LOCUS       sequence-415348-        8991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Premade plasma membrane marker, pUBQ10:PiP1;4:mTurquoise2:tHSP18.2M,
            for braiding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8991)
  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 8991)
  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: pKD013
FEATURES             Location/Qualifiers
     source          1..8991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1611..1613,1614..1616,1617..2330)
                     /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(1656..1677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1917..1936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2264..2285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2656..2680
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3981..4610
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5039..6112
                     /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      6178..6372
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6608..6630)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6716..6856
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6771..6790
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6871..6888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7042..7630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7122..7141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7718..8533)
                     /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(7718..8533)
                     /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     8441..8460
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    8966..8990
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacgaatta ccatggcacc tgctgcaacc tcgacgagtc agtaataaac ggcgtcaaag
       61 tggttgcagc cggcacacac gagtcgtgtt tatcaactca aagcacaaat acttttcctc
      121 aacctaaaaa taaggcaatt agccaaaaac aactttgcgt gtaaacaacg ctcaatacac
      181 gtgtcatttt attattagct attgcttcac cgccttagct ttctcgtgac ctagtcgtcc
      241 tcgtcttttc ttcttcttct tctataaaac aatacccaaa gagctcttct tcttcacaat
      301 tcagatttca atttctcaaa atcttaaaaa ctttctctca attctctcta ccgtgatcaa
      361 ggtaaatttc tgtgttcctt attctctcaa aatcttcgat tttgttttcg ttcgatccca
      421 atttcgtata tgttctttgg tttagattct gttaatctta gatcgaagac gattttctgg
      481 gtttgatcgt tagatatcat cttaattctc gattagggtt tcatagatat catccgattt
      541 gttcaaataa tttgagtttt gtcgaataat tactcttcga tttgtgattt ctatctagat
      601 ctggtgttag tttctagttt gtgcgatcga atttgtagat taatctgagt ttttctgatt
      661 aacaaacagt attcagtcga ctggtaccaa caggctcaac aatggaaggc aaagaagaag
      721 atgtacgagt gggagctaac aagttcccgg agaggcaacc catcggtaca tcggctcagt
      781 ccaccgacaa ggactacaaa gagccacctc ctgcgccact gttcgagccc ggcgagctca
      841 gctcatggtc tttctacaga gccggaatag ctgagtttat cgctactttc ttgtttctct
      901 acatcactgt tttgactgta atgggagtta agagagcacc aaacatgtgt gcttctgttg
      961 gaatccaagg tatcgcgtgg gcttttggtg gcatgatctt tgctcttgtc tactgtactg
     1021 ctggaatttc aggtggacac atcaaccctg ctgtaacatt cggtctgttc ttggctcgga
     1081 agttatctct gaccagagca gtgttctaca tgattatgca atgccttgga gccatctgtg
     1141 gtgccggagt cgtcaaaggt ttccagccaa cgccgtacca gactctcggt ggtggtgcta
     1201 acaccgttgc tcatggctac accaaaggtt ctggccttgg tgctgaaatc atcggaacat
     1261 tcgttctcgt ctacactgtc ttctccgcca ccgacgccaa gagaagcgcc cgtgactcac
     1321 acgtcccgat tttggcgccg ctcccaatcg gatttgcagt gttcttggta cacttggcaa
     1381 caataccgat caccgggacc ggaatcaacc cagctagaag tcttggagcc gcaattatct
     1441 acaacaagga ccactcttgg gatgaccatt ggattttctg ggttggacca ttcattggag
     1501 cagctctagc agcactatat caccagattg tcatcagagc gattcctttc aagagcaaga
     1561 gttcaggtgc tagcggtggc agcggtggca ctagtggtgg aggcggatcc atggtgagca
     1621 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1681 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1741 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1801 ccctgtcctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1861 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1921 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1981 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2041 acaactactt tagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     2101 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     2161 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2221 cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2281 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag ctgcgagctc
     2341 atatgaagat gaagatgaaa tatttggtgt gtcaaataaa tagcttgtgt gcttaagttt
     2401 gtgttttttt cttggcttgt tgtgttatga atttgtggct ttttctaata ttaaatgaat
     2461 gtaagatctc attataatga ataaacaaat gtttctataa tccattgtga atgttttgtt
     2521 ggatctcttc tgcagcatat aactactgta tgtgctatgg tatggactat ggaatatgat
     2581 taaagataag gaattcacta agtgacacat gaccatagga gaacgtatgt catatggcag
     2641 gtgctcgagc aattctgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta
     2701 ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg
     2761 tgcatgccaa ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg
     2821 ctatagtgca gtcggcttct gacgttcagt gcagccgtca tctgaaaacg acatgtcgca
     2881 caagtcctaa gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc
     2941 gtgttttagt cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa
     3001 caagagcgcc gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt
     3061 gaccaaccaa cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat
     3121 caccggcacc aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg
     3181 cgacgttgtg acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga
     3241 cattgccgag cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc
     3301 cgacaccacc acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt
     3361 cgagcgttcc ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg
     3421 cgtgaagttt ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct
     3481 gatcgaccag gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc
     3541 gaccctgtac cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg
     3601 cggtgccttc cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga
     3661 acgccaagag gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt
     3721 accgaagaga tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac
     3781 ctctcaaccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc
     3841 tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta
     3901 tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa
     3961 caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag
     4021 gcaagacgac catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc
     4081 tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc
     4141 aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg
     4201 gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg
     4261 cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg
     4321 ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac
     4381 aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg
     4441 aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct
     4501 acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg
     4561 cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg
     4621 taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag
     4681 cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt
     4741 tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac
     4801 cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat
     4861 aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag
     4921 gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg
     4981 gctgggttgc ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt
     5041 gagcggtcgc aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt
     5101 ggagaagttg aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc
     5161 cggtgaatcg tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc
     5221 agccggtgcg ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt
     5281 tccgatgctc tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt
     5341 ccgtctgtcg aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg
     5401 gcacgtagag gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt
     5461 actgatggcg gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga
     5521 caagcccggc cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc
     5581 cgatggcgga aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca
     5641 cgttgccatg cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg
     5701 tgaagccttg attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat
     5761 cgagatcgag ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt
     5821 gctgacggtt caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg
     5881 cctggcacgc cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga
     5941 acgcagtggc agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg
     6001 gtcaaatgac ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct
     6061 agtcatgcgc taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga
     6121 gcagatgcta gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggac tctttcctgt
     6181 ggatagcacg tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg
     6241 gaacccaaag ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa
     6301 aaaaggcgat ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct
     6361 ggcctgtgca taactgtctg gccagcgcac agccgaagag ctgcaaaaag cgcctaccct
     6421 tcggtcgctg cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg
     6481 ctcaaaaatg gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc
     6541 gccactcgac cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg
     6601 gtgaaaacct ctgacacatg cagctcccgg tgacggtcac agcttgtctg taagcggatg
     6661 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     6721 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     6781 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6841 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6901 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6961 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7441 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7681 ctcacgttaa gggattttgg tcatgcattc taggtgatta gaaaaactca tcgagcatca
     7741 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     7801 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     7861 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     7921 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     7981 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     8041 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgagt cgaaatacgc
     8101 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     8161 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     8221 ttttccctgg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     8281 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     8341 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     8401 catacaatcg gtagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     8461 catataaatc agcatccatg ttggaattta atcgcggcct tgagcaagac gtttcccgtt
     8521 gaatatggct cataacagaa cttattattt ccttcctctt ttctacagta tttaaagata
     8581 ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac
     8641 cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta
     8701 tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac
     8761 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg
     8821 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc
     8881 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
     8941 ggtcggggag ctgttggctg gctggtggca ggatatattg tggtgtaaac a
//
