LOCUS       sequence-317976-        8010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration in the chloroplast trnG_trnfM region: HS1-L, genome 
            position: 40216_ 41843, Genbank accession no. MH635409.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8010)
  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 8010)
  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: L1-Ck1-hs1-l-chlo
FEATURES             Location/Qualifiers
     source          1..8010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..128)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    214..354
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     269..288
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(369..386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(540..1128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(620..639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1216..2031)
                     /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(1216..2031)
                     /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     1939..1958
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2464..2488
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    4164..4188
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5489..6118
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6547..7620
                     /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      7686..7880
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca
       61 catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
      121 gctcccggtg acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
      181 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
      241 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
      301 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctca
      361 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      421 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      481 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      541 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      601 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      661 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      721 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      781 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      841 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      901 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      961 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1021 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1081 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1141 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1201 atgcattcta ggtgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1261 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1321 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1381 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1441 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     1501 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     1561 attcattcgt gattgcgcct gagcgagtcg aaatacgcga tcgctgttaa aaggacaatt
     1621 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     1681 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttccctggga tcgcagtggt
     1741 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1801 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1861 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcggt agattgtcgc
     1921 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1981 ggaatttaat cgcggccttg agcaagacgt ttcccgttga atatggctca taacagaact
     2041 tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct aattataaca
     2101 agacgaactc caattcactg ttccttgcat tctaaaacct taaataccag aaaacagctt
     2161 tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg attttgaaac
     2221 cgcggtgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg
     2281 agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa
     2341 catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg
     2401 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
     2461 tggtggcagg atatattgtg gtgtaaacat aacgaattgc tcttcaatga ggaggattgt
     2521 tagtgtagat aatttagaag atattattgg cgggcatgta tggttaggtt ctatttgtat
     2581 ttttggggga atctggcata ttttaacaaa accttttgca tgggctcgtc gtgctttagt
     2641 atggtctgga gaagcttact tatcttatag tttaggtgct attgctgttt ttggttttat
     2701 tgcttgttgt tttgtttggt tcaataatac tgcttatccg agtgagtttt atggtcctac
     2761 tggtccagaa gcttctcaag ctcaagcttt tactttttta gttagagatc aacgtcttgg
     2821 agctaatgta ggttcagctc aagggcctac tggattagga aaatatatta tgcgttcacc
     2881 tactggagaa attatttttg gtggagaaac aatgcgtttt tgggatcttc gtgctccatg
     2941 gttagaacca ttacgtggac caaacggttt agatctaagt aaattaaaaa aagatataca
     3001 accttggcaa gaacgtcgat ctgcagaata catgacacat gctccattag gatcattaaa
     3061 ttctgtaggt ggagtagcaa cagaaattaa tgcagtaaat tatgtttctc ctagaagttg
     3121 gttagcaaca tctcattttg ttttaggttt ctttttcttt gtagggcact tatggcatgc
     3181 tggtagagca cgtgctgctg cagctggttt tgaaaaagga attgatcgtg attttgaacc
     3241 agttctttct atgacacctc ttaattaatt attaacaagt aaaaaaacaa gtaaaaaaac
     3301 aaataaattg tttttttact tgttttttta cttgtttttt tacttaataa ttaacttgat
     3361 gatttaacta tttttttttt aattcaaaaa ggagagagag ggatttgaac cctcgataat
     3421 cttaaaaact atatcggttt tcaagaccga cgccataaac cactcggcca tctctcctat
     3481 agtaaacatt tttaatctaa taatttttta ttaaaaaatt ataaaaattt gatcaaattt
     3541 tttgtttttt tatcaactat ttgaaattat aagtaatttt ttgataattt tacaaaaaca
     3601 ggatttgatg gtaatatttt catatttatt aaaaacttgg agaatcacaa ctatgactat
     3661 agctttccaa tttgctgtgt ttgcactaat tgctatttca tttctcctag taattggtgt
     3721 tcccgtagta ctagcttctc ctgaaggttg gtcaagtaac aaaaatgttg ttttttcagg
     3781 tgcttcttta tggattggac tagttttttt agtaggtatt cttaattcgt ttatatctta
     3841 aaattttata gtaatctaaa atttatgaat ttaaaccttc cttggtttac attatattat
     3901 aaattctaaa tgcatttgaa acaagtgctt tagataaaaa aaaatgtttc caaggaggtt
     3961 taattaaaaa aaatataaaa aattctataa ataaataaaa aaaaatatat attatatata
     4021 taataagtat gtataattta tatacgcggg tatagtttaa tggtaaaatt cctccttgcc
     4081 aaggagaaga tgcgggttcg attcccgcta cccgccaatt gaagtaatta gcttagaaaa
     4141 aacgctagca tgaagagcaa ttctgacagg atatattggc gggtaaacct aagagaaaag
     4201 agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
     4261 tttgtatgtg catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc
     4321 ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcatc tgaaaacgac
     4381 atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt
     4441 cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac
     4501 gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac
     4561 caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc
     4621 gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta
     4681 cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac
     4741 ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag
     4801 ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt
     4861 gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag
     4921 gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc
     4981 cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg
     5041 catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc
     5101 aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc
     5161 gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt
     5221 ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc
     5281 ccgcgcacct ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc
     5341 tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt
     5401 gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag
     5461 taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg
     5521 cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc
     5581 cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg
     5641 ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa
     5701 ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc
     5761 tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga
     5821 catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg
     5881 aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga
     5941 ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg
     6001 cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg
     6061 cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt
     6121 taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc
     6181 gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg
     6241 gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa
     6301 ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc
     6361 aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga
     6421 acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg
     6481 cgtaagcggc tgggttgcct gccggccctg caatggcact ggaaccccca agcccgagga
     6541 atcggcgtga gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat
     6601 gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa
     6661 gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa
     6721 ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat
     6781 tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg
     6841 gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt
     6901 ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac
     6961 gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg
     7021 aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc
     7081 cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac
     7141 accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta
     7201 tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg
     7261 gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac
     7321 ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt
     7381 ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg
     7441 atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag
     7501 ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc
     7561 ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa
     7621 tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggactc
     7681 tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg
     7741 tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata
     7801 aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa
     7861 acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagtgct gcaaaaagcg
     7921 cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc
     7981 gctggccgct caaaaatggc tggcctacgg
//
