LOCUS       sequence-274974-        6911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use when making odd level constructs (L3,...) if pCsB position 
            (position 2) has no construct assigned.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6911)
  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 6911)
  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: pCsB_spacer
FEATURES             Location/Qualifiers
     source          1..6911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1563..1587
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    1835..1859
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3160..3789
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4218..5291
                     /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      5357..5551
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5787..5809)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5895..6035
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5950..5969
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6050..6067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6221..6809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6301..6320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6897..6911,1..777))
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
       61 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
      121 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
      181 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcaccagc
      241 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
      301 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
      361 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
      421 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
      481 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
      541 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
      601 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
      661 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
      721 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
      781 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
      841 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
      901 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
      961 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     1021 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     1081 cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg aacagaactt
     1141 attatttcct tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa
     1201 gacgaactcc aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt
     1261 ttcaaagttg ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc
     1321 gcggtgatca caggcagcaa cgctctgtca tcgttacaat caacatgcta ccctccgcga
     1381 gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag catcggtaac
     1441 atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg gactgatggg
     1501 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct
     1561 ggtggcagga tatattgtgg tgtaaacata acggatccgg tctcatacta atggtagaca
     1621 ctttattagt tagatccgct gttattctgc acgattgcgc ttgccggtgg aaatttaagg
     1681 acgagtgtag tgcactcacc cttagccttc agcgtgacaa gattagggta gcaccggaaa
     1741 cagcgatttt acggacagat agtacagtct gagactcagt cactggtccc taaacgggag
     1801 ccggtacgtc ggcggtaaat gtgagaccgg atcctgacag gatatattgg cgggtaaacc
     1861 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
     1921 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt
     1981 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtcat
     2041 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
     2101 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
     2161 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
     2221 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
     2281 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
     2341 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
     2401 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
     2461 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
     2521 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
     2581 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
     2641 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
     2701 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
     2761 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
     2821 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
     2881 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
     2941 gttcgagccg cccgcgcacc tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
     3001 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     3061 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     3121 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     3181 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
     3241 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
     3301 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
     3361 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
     3421 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
     3481 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
     3541 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
     3601 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
     3661 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
     3721 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
     3781 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     3841 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     3901 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     3961 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     4021 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     4081 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     4141 ggttggccag gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc
     4201 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
     4261 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
     4321 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
     4381 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg
     4441 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
     4501 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
     4561 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
     4621 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
     4681 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
     4741 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
     4801 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
     4861 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
     4921 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
     4981 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
     5041 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
     5101 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
     5161 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
     5221 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
     5281 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
     5341 gaaaaggact ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
     5401 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
     5461 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
     5521 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagtgc
     5581 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc
     5641 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc
     5701 gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc
     5761 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccggt gacggtcaca
     5821 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     5881 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     5941 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     6001 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc atccgcttcc tcgctcactg
     6061 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6121 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6181 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6241 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6301 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6361 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6421 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6481 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6541 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6601 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6661 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6721 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6781 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6841 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct aggtgattat
     6901 ttgccgacta c
//
