LOCUS       sequence-331900-        7906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration in the chloroplast rbcL_trnR region: HS2-L, genome 
            position: 56139_57662, Genbank accession no. MH635409.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7906)
  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 7906)
  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-hs2-l-chlo
FEATURES             Location/Qualifiers
     source          1..7906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      361..555
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(791..813)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    899..1039
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     954..973
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1054..1071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1225..1813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1305..1324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1901..2716)
                     /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(1901..2716)
                     /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     2624..2643
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    3149..3173
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    4745..4769
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6070..6699
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(7128..7906,1..295)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta
       61 ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta
      121 cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat
      181 cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat
      241 cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac
      301 ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gactctttcc
      361 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat
      421 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga
      481 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg
      541 cctggcctgt gcataactgt ctggccagcg cacagccgaa gtgctgcaaa aagcgcctac
      601 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg
      661 ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc
      721 gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg
      781 acggtgaaaa cctctgacac atgcagctcc cggtgacggt cacagcttgt ctgtaagcgg
      841 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
      901 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
      961 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     1021 gagaaaatac cgcatcaggc gctcatccgc ttcctcgctc actgactcgc tgcgctcggt
     1081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     1681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1861 aaactcacgt taagggattt tggtcatgca ttctaggtga ttagaaaaac tcatcgagca
     1921 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1981 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2041 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2101 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     2161 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     2221 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agtcgaaata
     2281 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     2341 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     2401 ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     2461 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     2521 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     2581 tcccatacaa tcggtagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     2641 acccatataa atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc
     2701 gttgaatatg gctcataaca gaacttatta tttccttcct cttttctaca gtatttaaag
     2761 ataccccaag aagctaatta taacaagacg aactccaatt cactgttcct tgcattctaa
     2821 aaccttaaat accagaaaac agctttttca aagttgtttt caaagttggc gtataacata
     2881 gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg cagcaacgct ctgtcatcgt
     2941 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
     3001 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
     3061 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
     3121 gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacataacga
     3181 attgctcttc aatgaggagc agcatttaga atgactcctc aacctggagt tccagcagaa
     3241 gaagcaggag cagcagttgc tgctgagtct tcaactggta catggactac agtttggact
     3301 gatggtctta ctaaccttga tcgttataaa ggtcgatgct atgatattga ccctgttcct
     3361 ggagaagata atcaatatat tgcttatgta gcttatcctt tagatttatt tgaagaaggt
     3421 tctgttacaa atatgtttac ttcaattgta ggtaatgtat ttgggtttaa agctttaaga
     3481 gcgttacgtc ttgaagattt aagaattcct ccagcttaca caaaaacttt ccaaggtcct
     3541 cctcatggta ttcaagttga gagagataaa ttaaacaaat atggtcgtcc tttattagga
     3601 tgtactatta aaccaaaatt aggtttatct gctaaaaact atggtagagc tgtatatgaa
     3661 tgtcttcgtg gtggacttga ttttactaaa gatgatgaaa acgtaaattc tcaaccattt
     3721 atgcgttgga gagatcgttt cttatttgta gcagaagcta tttttaaatc tcaagcagaa
     3781 actggagaaa tcaaaggaca ttatttaaat gctactgcag gtacatgtga agaaatgcta
     3841 aaaagagcag catgtgctag agagttaggt gtaccaattg ttatgcacga ctacttaact
     3901 ggtggtttca ctgcaaatac tactctagct ttttattgcc gtgacaatgg tttacttctt
     3961 catattcacc gtgcaatgca tgcagttatt gatagacaaa aaaatcatgg tatccacttc
     4021 cgtgtattag caaaagcttt acgtatgtct ggtggagatc atattcacgc tggtactgtt
     4081 gtaggtaaac ttgaaggaga tcgtaacgta actttaggtt tcgtagattt acttcgtgat
     4141 gactatattg aaaaagatag aagtcgtggt atttatttca cacaagattg ggtttcttta
     4201 cctggtgttt tccctgtagc atctggtggg atccatgttt ggcatatgcc tgctttaact
     4261 gaaatttttg gcgatgactc tgttttacaa tttggtggtg gaactttagg tcatccttgg
     4321 ggtaacgcac ctggtgcagt tgctaaccga gtttcgttag aagcttgtgt acaagcacgt
     4381 aatgaaggtc gtgaccttgc tcgtgaagga aatgaaatta ttcgtgaagc ttgcaaatgg
     4441 agtcctgagt tatctgctgc ttgtgaagtt tggaaagaaa ttaaatttga atttgatatt
     4501 attgatactt tgtaaaatga aagtagatat actttgtaaa ataaagtaga tattttatct
     4561 taaaagtttt gtaattttct tttttttatc tgatatttca gataaaaaaa agaaaattac
     4621 aaaactttta aatgggtttg tagctcagtg gattagagct catggttccg gatcatgaag
     4681 tcaagggttc gaatcccttc taaccctttt tttattttga aaacgctagc atgaagagca
     4741 attctgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc
     4801 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac
     4861 cacagggttc ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag
     4921 tcggcttctg acgttcagtg cagccgtcat ctgaaaacga catgtcgcac aagtcctaag
     4981 ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc
     5041 gcataaagta gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg
     5101 ccgctggcct gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac
     5161 gggccgaact gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca
     5221 ggcgcgaccg cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga
     5281 cagtgaccag gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc
     5341 gcatccagga ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca
     5401 cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc
     5461 taatcatcga ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg
     5521 gcccccgccc taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg
     5581 aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc
     5641 gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc
     5701 gtgaggacgc attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg
     5761 aacaagcatg aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat
     5821 cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacc tctcaaccgt
     5881 gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag
     5941 cttggccgct gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa
     6001 cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa
     6061 ggggaacgca tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc
     6121 atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat
     6181 tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc
     6241 gttgtcggca tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac
     6301 ttcgtagtga tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca
     6361 gccgacttcg tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac
     6421 ctggtggagc tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt
     6481 gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc
     6541 gggtacgagc tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact
     6601 gccgccgccg gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc
     6661 caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa
     6721 tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg
     6781 caacgttggc cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg
     6841 cggaggatca caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc
     6901 tgctatctga atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga
     6961 tgaattttag cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc
     7021 gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgcc
     7081 tgccggccct gcaatggcac tggaaccccc aagcccgagg aatcggcgtg agcggtcgca
     7141 aaccatccgg cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga
     7201 aggccgcgca ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt
     7261 ggcaagcggc cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc
     7321 cgtcgattag gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct
     7381 atgacgtggg cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga
     7441 agcgtgaccg acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg
     7501 tttccgcagg gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg
     7561 tttcccatct aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc
     7621 gcgtgttccg tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa
     7681 agcagaaaga cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc
     7741 agcgtacgaa gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga
     7801 ttagccgcta caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc
     7861 tagctgattg gatgtaccgc gagatcacag aaggcaagaa cccgga
//
