LOCUS       sequence-331912-        8006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration in the chloroplast rbcL_trnR region: HS2-R, genome 
            position: 57681- 59304, Genbank accession no. MH635409.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8006)
  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 8006)
  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-Ck4-hs2-r-chlo
FEATURES             Location/Qualifiers
     source          1..8006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    433..457
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    2129..2153
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3454..4083
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4512..5585
                     /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      5651..5845
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6081..6103)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6189..6329
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6244..6263
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6344..6361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6515..7103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6595..6614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7191..8006)
                     /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(7191..8006)
                     /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     7914..7933
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aacagaactt attatttcct tcctcttttc tacagtattt aaagataccc caagaagcta
       61 attataacaa gacgaactcc aattcactgt tccttgcatt ctaaaacctt aaataccaga
      121 aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa catagtatcg acggagccga
      181 ttttgaaacc gcggtgatca caggcagcaa cgctctgtca tcgttacaat caacatgcta
      241 ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag
      301 catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg
      361 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
      421 ttggctggct ggtggcagga tatattgtgg tgtaaacata acgaattgct cttcacagag
      481 gagattgttc tattattaaa ttacttaaat tttaaatttt tcatatatgt tttaaaaata
      541 attatatata aaaatatgga acattttttt tataattatt ttctcttaaa attttaaaaa
      601 atttataaaa aaagaaataa agaggatttt tttatgtctt taatgaattg gtttgaagat
      661 aaacgaagat ttggtggatt aattggcgct tttattgaaa aagctactaa aggatatatt
      721 tttagtgaaa gagaaaaaga tcgatatatt aaaattgaca ctactaaggg attatggact
      781 agatgtgaca attgtgaaaa tatgttatat gttagatttt tgagacaaaa taaacgaatt
      841 tgcgaagaat gcggatatca tttgcaaatg agtagtacag aaagaattga acttttaatt
      901 gatcgcggga cttggtatcc aatggatgaa gatatgactg ctcgagatgt tcttaaattt
      961 tctgatgaag attcttataa aaatcgaatt gctttttatc aaaaaagaac tggtttaact
     1021 gatgcaattc aaacaggcat aggtcaatta aacggtattc ctattgcatt gggagtaatg
     1081 gattttcaat ttatgggcgg tagtatggga tctgtagtag gtgaaaaaat tactcgtctt
     1141 attgaatatg ctactagagc atcaatgcca ctaattatag tatgttcttc tggtggagca
     1201 cgcatgcaag aaggaacatt aagtttaatg caaatggcta aaatttcttc ggttttgcaa
     1261 attcatcaag cccaaaaaag attactttat atagctattc ttacctatcc tacaacagga
     1321 ggagttactg caagttttgg tatgttaggg gatattatta ttgctgagcc aaaagcttat
     1381 attgcatttg caggaaaaag agttattgaa caaactttac gacaaaaaat accagatggt
     1441 tttcaagttg cagaatcatt atttgatcat ggtttacttg atttaattgt tccacgaaat
     1501 cttttaaaag gtgttttaag tgaaattttt gaattatata atgctgctcc gtgtaaaaaa
     1561 tttcaaaatt ctttttttta ataattttgt tagattttta gtagtttttt ttaattcaaa
     1621 ttttttataa atttttatat tttgttattg ttatttatta aaataagata taatttttat
     1681 ttagtttagt tttaaataaa atatagtata tattatattt attttttttt ttttattgta
     1741 ttttaaggta ttttttatga cagcttctta tttaccttct atttttgttc ctttagttgg
     1801 attaattttt cctgctatta ctatggcttc attatttata tatattgaac aagatgaaat
     1861 tttataaatt aattggagat taaaattttt ttttactttt tacaaaatat atatatgtta
     1921 tagatcaatt tttttttgat atatatttta taatactttg tgaaaagtaa aaaatttttt
     1981 ttgaatttat tgttattata atattgatta ttttttaata gaaaaataat caatattttt
     2041 tttatcaaca ttattattat tcaactaatt taggagacat tcttttgtta tgaatttaca
     2101 agtggaccgc taggttgaag agcaattctg acaggatata ttggcgggta aacctaagag
     2161 aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc
     2221 gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggatcaaagt actttgatcc
     2281 aacccctccg ctgctatagt gcagtcggct tctgacgttc agtgcagccg tcatctgaaa
     2341 acgacatgtc gcacaagtcc taagttacgc gacaggctgc cgccctgccc ttttcctggc
     2401 gttttcttgt cgcgtgtttt agtcgcataa agtagaatac ttgcgactag aaccggagac
     2461 attacgccat gaacaagagc gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg
     2521 acgaccagga cttgaccaac caacgggccg aactgcacgc ggccggctgc accaagctgt
     2581 tttccgagaa gatcaccggc accaggcgcg accgcccgga gctggccagg atgcttgacc
     2641 acctacgccc tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc
     2701 gcgacctact ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg
     2761 cagagccgtg ggccgacacc accacgccgg ccggccgcat ggtgttgacc gtgttcgccg
     2821 gcattgccga gttcgagcgt tccctaatca tcgaccgcac ccggagcggg cgcgaggccg
     2881 ccaaggcccg aggcgtgaag tttggccccc gccctaccct caccccggca cagatcgcgc
     2941 acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa agaggcggct gcactgcttg
     3001 gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg
     3061 aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac gccctggcgg
     3121 ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg caccaggacg gccaggacga
     3181 accgtttttc attaccgaag agatcgaggc ggagatgatc gcggccgggt acgtgttcga
     3241 gccgcccgcg cacctctcaa ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc
     3301 caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa
     3361 aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg
     3421 atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag
     3481 aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc
     3541 ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc
     3601 gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac
     3661 gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac
     3721 ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct
     3781 tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg
     3841 gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc
     3901 ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg
     3961 cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc
     4021 gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt
     4081 tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc
     4141 cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga
     4201 agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac
     4261 gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa
     4321 tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat
     4381 caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg
     4441 ccaggcgtaa gcggctgggt tgcctgccgg ccctgcaatg gcactggaac ccccaagccc
     4501 gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg
     4561 gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg
     4621 cagaagcacg ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc
     4681 ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac
     4741 cagatttttt cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg
     4801 acgtggccgt tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg
     4861 agcttccaga cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg
     4921 attacgacct ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg
     4981 aagggaaggg agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt
     5041 tctgccggcg agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt
     5101 taaacaccac gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga
     5161 cggtatccga gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc
     5221 ggccggagta catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca
     5281 agaacccgga cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc
     5341 gttttctcta ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca
     5401 agacgatcta cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc
     5461 gcaagctgat cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg
     5521 ctggcccgat cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt
     5581 cctaatgtac ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag
     5641 gactctttcc tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga
     5701 acccgtacat tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg
     5761 atataaaaga gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc
     5821 ttaaaacccg cctggcctgt gcataactgt ctggccagcg cacagccgaa gtgctgcaaa
     5881 aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg
     5941 cggccgctgg ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac
     6001 aagccgcgcc gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt
     6061 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggtgacggt cacagcttgt
     6121 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6181 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
     6241 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     6301 gatgcgtaag gagaaaatac cgcatcaggc gctcatccgc ttcctcgctc actgactcgc
     6361 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6421 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6481 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6541 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6601 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6661 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6721 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6781 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6841 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6901 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6961 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7021 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7081 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7141 agtggaacga aaactcacgt taagggattt tggtcatgca ttctaggtga ttagaaaaac
     7201 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     7261 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     7321 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     7381 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     7441 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     7501 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     7561 agtcgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     7621 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     7681 acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta
     7741 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     7801 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     7861 gcatcgggct tcccatacaa tcggtagatt gtcgcacctg attgcccgac attatcgcga
     7921 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa
     7981 gacgtttccc gttgaatatg gctcat
//
