LOCUS       sequence-274964-        6582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use when making even level constructs (L2,...) if pCk1 position 
            (position 1) has no construct assigned.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  AUTHORS   Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M, 
            Markel K, Delmans M, West A, Patron NJ, Haseloff J
  TITLE     Systematic tools for reprogramming plant gene expression in a simple
            model, Marchantia polymorpha.
  JOURNAL   ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511.
  PUBMED    32163700
REFERENCE   2  (bases 1 to 6582)
  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: "ACS Synth 
            Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511."
COMMENT     Sequence Label: pCk1_spacer
FEATURES             Location/Qualifiers
     source          1..6582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     153..172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     406..423
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    438..578
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(504..523)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     664..686
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      922..1116
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(1182..2255)
                     /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"
     CDS             complement(2684..3313)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    4614..4638
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    4886..4910
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             5343..6158
                     /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             5343..6158
                     /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     complement(5416..5435)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(6246..6582,1..252)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
       61 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      121 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      181 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      241 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      301 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      361 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      421 gaggaagcgg atgagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      481 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
      541 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
      601 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
      661 tcaccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagg
      721 gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc gcgccctggt
      781 agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat aggccgacgc
      841 gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg cagcacttcg
      901 gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt
      961 tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg
     1021 tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg
     1081 tacggctttg ggttcccaat gtacgtgcta tccacaggaa agagtccttt tcgacctttt
     1141 tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct
     1201 tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc
     1261 tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag
     1321 aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg
     1381 gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga
     1441 tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc
     1501 tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc
     1561 tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg
     1621 ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt
     1681 tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc
     1741 gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc
     1801 atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc
     1861 atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc
     1921 tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga
     1981 ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg ctcgtcgccc
     2041 ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga ttctttgcgg
     2101 attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc gatgcgttgc
     2161 cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag cgccgcgccg
     2221 atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc ttgggggttc
     2281 cagtgccatt gcagggccgg caggcaaccc agccgcttac gcctggccaa ccgcccgttc
     2341 ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt tccatgccgc
     2401 ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta ctctggtagc
     2461 tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac cgcgtacatc
     2521 ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat ggctggcgtg
     2581 tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg gccggcactt
     2641 agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg agttttgatt
     2701 taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt tctgattcaa
     2761 gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg atacgggact
     2821 caagaatggg cagctcgtac ccggccagcg cctcggcaac ctcaccgccg atgcgcgtgc
     2881 ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct tccatccgtg acctcaatgc
     2941 gctgcttaac cagctccacc aggtcggcgg tggcccatat gtcgtaaggg cttggctgca
     3001 ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc gccgcctggg
     3061 gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg cggtcaatcg
     3121 tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc gcccaatcgc
     3181 gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg agttgcaggg
     3241 cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg ttaagtacag
     3301 cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca tcatatacgc
     3361 agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta gacggcggcg
     3421 ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat cagacaaacc
     3481 ggccaggatt tcatgcagcc gcacggttga gaggtgcgcg ggcggctcga acacgtaccc
     3541 ggccgcgatc atctccgcct cgatctcttc ggtaatgaaa aacggttcgt cctggccgtc
     3601 ctggtgcggt ttcatgcttg ttcctcttgg cgttcattct cggcggccgc cagggcgtcg
     3661 gcctcggtca atgcgtcctc acggaaggca ccgcgccgcc tggcctcggt gggcgtcact
     3721 tcctcgctgc gctcaagtgc gcggtacagg gtcgagcgat gcacgccaag cagtgcagcc
     3781 gcctctttca cggtgcggcc ttcctggtcg atcagctcgc gggcgtgcgc gatctgtgcc
     3841 ggggtgaggg tagggcgggg gccaaacttc acgcctcggg ccttggcggc ctcgcgcccg
     3901 ctccgggtgc ggtcgatgat tagggaacgc tcgaactcgg caatgccggc gaacacggtc
     3961 aacaccatgc ggccggccgg cgtggtggtg tcggcccacg gctctgccag gctacgcagg
     4021 cccgcgccgg cctcctggat gcgctcggca atgtccagta ggtcgcgggt gctgcgggcc
     4081 aggcggtcta gcctggtcac tgtcacaacg tcgccagggc gtaggtggtc aagcatcctg
     4141 gccagctccg ggcggtcgcg cctggtgccg gtgatcttct cggaaaacag cttggtgcag
     4201 ccggccgcgt gcagttcggc ccgttggttg gtcaagtcct ggtcgtcggt gctgacgcgg
     4261 gcatagccca gcaggccagc ggcggcgctc ttgttcatgg cgtaatgtct ccggttctag
     4321 tcgcaagtat tctactttat gcgactaaaa cacgcgacaa gaaaacgcca ggaaaagggc
     4381 agggcggcag cctgtcgcgt aacttaggac ttgtgcgaca tgtcgttttc agatgacggc
     4441 tgcactgaac gtcagaagcc gactgcacta tagcagcgga ggggttggat caaagtactt
     4501 tgatcccgag gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct
     4561 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
     4621 cgccaatata tcctgtcaga attgctcttc atgctagcga ttgccatgtc gaagtccgtt
     4681 ataagggtca ttctaccatg cacgccatga ttctgaaagt actgacgagc tgcaagaacg
     4741 agatgcacac ttctcatcac cggggcacag acctcaggtg ggacgttggc caactcaggt
     4801 ccgtgtcgga agaacccaac gaacatgtaa ctctcactac gcgagttgtc ctgctcagac
     4861 tcctcattga agagcaattc gttatgttta caccacaata tatcctgcca ccagccagcc
     4921 aacagctccc cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag
     4981 tccgggacgg cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga
     5041 tgctattcgg aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag
     5101 ggtagcatgt tgattgtaac gatgacagag cgttgctgcc tgtgatcacc gcggtttcaa
     5161 aatcggctcc gtcgatacta tgttatacgc caactttgaa aacaactttg aaaaagctgt
     5221 tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg tcttgttata
     5281 attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata ataagttctg
     5341 ttatgagcca tattcaacgg gaaacgtctt gctcaaggcc gcgattaaat tccaacatgg
     5401 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     5461 tctaccgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     5521 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     5581 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     5641 cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     5701 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     5761 cttttaacag cgatcgcgta tttcgactcg ctcaggcgca atcacgaatg aataacggtt
     5821 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     5881 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     5941 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     6001 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     6061 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     6121 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc acctagaatg catgaccaaa
     6181 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6241 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6301 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6361 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6421 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6481 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6541 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca ca
//
