LOCUS       sequence-331903-        7738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L1 in position 2, for spectinomycin in chloroplast transformation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7738)
  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 7738)
  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-Ck2-smR-chlo
FEATURES             Location/Qualifiers
     source          1..7738
                     /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    1861..1885
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3186..3815
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4244..5317
                     /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      5383..5577
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5813..5835)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5921..6061
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5976..5995
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6076..6093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6247..6835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6327..6346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6923..7738)
                     /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(6923..7738)
                     /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     7646..7665
                     /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 cttcagcaag
      481 gagcgataag cttcgaatat agatcttctt tcttatttca atgatattat tatttcaaag
      541 ataagagata ttcaaagata agagataaga agaagtcaaa atttgatttt tttttggaaa
      601 aaaaaaatca aaaagatata gtaacattag caagaagaga aacaagttct atttctacaa
      661 ttttaaacaa atacaaaatc aaaatagaat actcaatcat gaataaatgc aagaaaataa
      721 cctctccttc tttttctata atgtaaacaa aaaagtctat gtaagtaaaa tactagtaaa
      781 taaataaaaa gaaaaaaaga aaggagcaat agcaccctct tgatagaaca agaaaatgat
      841 tattgctcct ttcttttcaa aacctcctat agactaggcc aggatcctgc agaagcttat
      901 ttgccaacta ccttagtgat ctcgcctttc acgtagtgga caaattcttc caactgatct
      961 gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg
     1021 acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc
     1081 gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc
     1141 tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca
     1201 aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca
     1261 acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc
     1321 tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta
     1381 gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg
     1441 agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc
     1501 cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc
     1561 ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga
     1621 tggcgctcga tgacgccaac tacctctgat agttgagttg atacttcggc gataaccgct
     1681 tcacgagcca ttggatccct ccctacaact gtatccaagc gcttcaaatt cgcccggagt
     1741 tcgctcccag aaatatagcc atccctgccc cctcacgtca atcccacgag cctcttatcc
     1801 attctcattg aacgacggcg ggggagcaaa tccaactagc gctatactga agagcaattc
     1861 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
     1921 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
     1981 gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg
     2041 cttctgacgt tcagtgcagc cgtcatctga aaacgacatg tcgcacaagt cctaagttac
     2101 gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat
     2161 aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc
     2221 tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc
     2281 cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg
     2341 cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt
     2401 gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat
     2461 ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc
     2521 ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat
     2581 catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc
     2641 ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg
     2701 ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc
     2761 acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga
     2821 ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca
     2881 agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag
     2941 gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacctctc aaccgtgcgg
     3001 ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg
     3061 gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc
     3121 ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg
     3181 aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg
     3241 caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg
     3301 atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg
     3361 tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg
     3421 tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg
     3481 acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg
     3541 tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg
     3601 tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt
     3661 acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg
     3721 ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg
     3781 cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag
     3841 caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac
     3901 gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga
     3961 ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct
     4021 atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa
     4081 ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg
     4141 aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc
     4201 ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc
     4261 atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc
     4321 cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca
     4381 agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc
     4441 gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga
     4501 cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg
     4561 tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc
     4621 cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc
     4681 ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt
     4741 gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca
     4801 gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg
     4861 tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag
     4921 ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc
     4981 tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc
     5041 cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc
     5101 cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc
     5161 cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc
     5221 ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg
     5281 caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca
     5341 aattgcccta gcaggggaaa aaggtcgaaa aggactcttt cctgtggata gcacgtacat
     5401 tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta
     5461 cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc
     5521 cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact
     5581 gtctggccag cgcacagccg aagtgctgca aaaagcgcct acccttcggt cgctgcgctc
     5641 cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg
     5701 cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg
     5761 gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5821 acatgcagct cccggtgacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5881 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     5941 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     6001 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6061 gcgctcatcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6121 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6181 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6661 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6901 tttggtcatg cattctaggt gattagaaaa actcatcgag catcaaatga aactgcaatt
     6961 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     7021 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     7081 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     7141 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     7201 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     7261 aaccgttatt cattcgtgat tgcgcctgag cgagtcgaaa tacgcgatcg ctgttaaaag
     7321 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     7381 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg
     7441 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     7501 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     7561 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcggtaga
     7621 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     7681 ccatgttgga atttaatcgc ggccttgagc aagacgtttc ccgttgaata tggctcat
//
