LOCUS       sequence-341232-        8158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration in the chloroplast trnG_trnfM region: HS1-R, genome 
            position: 41847_ 43622, Genbank accession no. MH635409 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8158)
  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 8158)
  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-hs1-r-chlo
FEATURES             Location/Qualifiers
     source          1..8158
                     /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"
     RBS             2243..2251
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    2281..2305
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3606..4235
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4664..5737
                     /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      5803..5997
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6233..6255)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6341..6481
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6396..6415
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6496..6513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6667..7255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6747..6766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7343..8158)
                     /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(7343..8158)
                     /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     8066..8085
                     /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 gagtaagtaa aaatattttt ttttgttttt tggcggagac aggatttgaa cctatgacct
      541 caaggttatg agccttgcga gctaccagac tgctctactc cgcgttataa ttaacgaaaa
      601 aagtatactc tataaaaata gaattaagca agatatattt taaagaaccc cgcctcttat
      661 aagaggcggg aaagaaactc aaaatgttta ccaactagat tttgttactc caggcaataa
      721 acatgcatga gccatttcac gaagtaaatg tctagataaa ccaaaatcgc ggtaatttgc
      781 tttaggtctt ccagtcaaaa aacaacgacg atgaagacga gtcggggcac tattacgtgg
      841 taaagattgt aatttttttt gaaattccca tttctcatct aatgatgaag tttccgtaat
      901 tttctttttt aaagaattac gtaaagtttt gtattttttt tctaaatttt gtcttttttt
      961 ttctctttga ataagactct tttttgccat aataatttaa ttagtaagat attctttttc
     1021 gataaatttt tattaaatta tatttttagt aaaattggaa atttactaca aatataattt
     1081 aaccaaattt accagacgta gaagcaatta aaaaagcagc ataagtaaat atataaccta
     1141 cagaaaaatg agctaatcca actaatcttg cttgaacaat agaaagagct actggtttat
     1201 ctttccagcg aactaaattc gctaaaggag tacgttcgtg agcccaagct aaagtttcaa
     1261 taagttcttg ccaatatcca cgccatgata tcagaaacat aaatccagta gcccaaacta
     1321 aatgaccaaa taaaaacatc catgcccaaa cagacaaact gttcatacca aaaggattat
     1381 atccattaat caattgtgaa gaatttaacc ataaataatc tcttaaccag cccattaaat
     1441 atgtagaaga ttcattaaat tgtgctgcat ttccttgcca taatgtaata tgtttccaat
     1501 gccaataaaa agtaacccat ccaatagtat ttaacatcca aaaaactgct aaataaaaag
     1561 catcccaagc agaaatatca caagtaccac ctcgtccagg accatcacaa ggaaaactat
     1621 aaccaaattc ttttttatct ggcattaatt tagatcctcg tgcatctaaa gcacctttca
     1681 ctaaaattaa tgtagtagta tgtaaaccta aagcaatagc gtgatgtact aaaaagtctc
     1741 caggaccaat tgttaagaaa agtgaattac tattattatt gatagcatct aaccacccag
     1801 gtaaccatat actttgacca gcattaaatg ctggattatt tgttgatgat aaaagtacat
     1861 caaaaccata taaagcttta ccatgagcag attgtatcca ttgagcaaaa ataggttcaa
     1921 ttaaaatttg tttttcagga gtaccaaaag caagcataac atcattatga acgtaaagac
     1981 ctaaagtatg aaaccctaga aataaactag cccaacttaa atgggatatt atagcttctt
     2041 tatgttctaa cattctagct aatacattat ctttattttg ttccggatta taatctctaa
     2101 taaagaaaat agctccgtga gcaaaggcac ctgtcataat aaaaccagca atatattgat
     2161 gatgagtata taatgccgct tgcgttgtaa aatcttgtgc aagaaacgca tacggaggta
     2221 aagaatacat atgttgagct actaaggagg taataactcc gctaggttga agagcaattc
     2281 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
     2341 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
     2401 gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg
     2461 cttctgacgt tcagtgcagc cgtcatctga aaacgacatg tcgcacaagt cctaagttac
     2521 gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat
     2581 aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc
     2641 tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc
     2701 cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg
     2761 cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt
     2821 gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat
     2881 ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc
     2941 ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat
     3001 catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc
     3061 ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg
     3121 ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc
     3181 acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga
     3241 ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca
     3301 agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag
     3361 gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacctctc aaccgtgcgg
     3421 ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg
     3481 gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc
     3541 ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg
     3601 aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg
     3661 caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg
     3721 atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg
     3781 tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg
     3841 tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg
     3901 acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg
     3961 tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg
     4021 tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt
     4081 acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg
     4141 ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg
     4201 cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag
     4261 caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac
     4321 gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga
     4381 ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct
     4441 atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa
     4501 ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg
     4561 aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc
     4621 ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc
     4681 atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc
     4741 cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca
     4801 agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc
     4861 gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga
     4921 cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg
     4981 tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc
     5041 cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc
     5101 ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt
     5161 gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca
     5221 gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg
     5281 tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag
     5341 ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc
     5401 tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc
     5461 cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc
     5521 cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc
     5581 cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc
     5641 ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg
     5701 caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca
     5761 aattgcccta gcaggggaaa aaggtcgaaa aggactcttt cctgtggata gcacgtacat
     5821 tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta
     5881 cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc
     5941 cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact
     6001 gtctggccag cgcacagccg aagtgctgca aaaagcgcct acccttcggt cgctgcgctc
     6061 cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg
     6121 cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg
     6181 gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     6241 acatgcagct cccggtgacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     6301 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     6361 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     6421 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6481 gcgctcatcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7081 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7321 tttggtcatg cattctaggt gattagaaaa actcatcgag catcaaatga aactgcaatt
     7381 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     7441 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     7501 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     7561 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     7621 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     7681 aaccgttatt cattcgtgat tgcgcctgag cgagtcgaaa tacgcgatcg ctgttaaaag
     7741 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     7801 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg
     7861 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     7921 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     7981 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcggtaga
     8041 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     8101 ccatgttgga atttaatcgc ggccttgagc aagacgtttc ccgttgaata tggctcat
//
