LOCUS       sequence-274971-        6911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use when making odd level constructs (L3,...) if pCsA position 
            (position 1) 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: pCsA_spacer
FEATURES             Location/Qualifiers
     source          1..6911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(73..864)
                     /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"
     misc_feature    1650..1674
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    1922..1946
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3247..3876
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4305..5378
                     /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      5444..5638
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5874..5896)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5982..6122
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6037..6056
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6137..6154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6308..6896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6388..6407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
       61 cattctaggt gattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
      121 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
      181 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
      241 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
      301 aagcactaca tttcgctcat caccagccca gtcgggcggc gagttccata gcgttaaggt
      361 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
      421 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
      481 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
      541 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
      601 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
      661 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
      721 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
      781 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
      841 ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac tgccctgctg
      901 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
      961 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac
     1021 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     1081 catacgctac ttgcattaca gcttacgaac cgaacaggct tatgtccact gggttcgtgc
     1141 cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggtagcagcg aagtcgaggc
     1201 atttctgtcc tggctggaac agaacttatt atttccttcc tcttttctac agtatttaaa
     1261 gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc ttgcattcta
     1321 aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg cgtataacat
     1381 agtatcgacg gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg
     1441 ttacaatcaa catgctaccc tccgcgagat catccgtgtt tcaaacccgg cagcttagtt
     1501 gccgttcttc cgaatagcat cggtaacatg agcaaagtct gccgccttac aacggctctc
     1561 ccgctgacgc cgtcccggac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc
     1621 tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacataacg
     1681 gatccggtct caggagaatg gtagacactt tattagttag atccgctgtt attctgcacg
     1741 attgcgcttg ccggtggaaa tttaaggacg agtgtagtgc actcaccctt agccttcagc
     1801 gtgacaagat tagggtagca ccggaaacag cgattttacg gacagatagt acagtctgag
     1861 actcagtcac tggtccctaa acgggagccg gtacgtcggc ggtatacttg agaccggatc
     1921 ctgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga
     1981 tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac
     2041 agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat agtgcagtcg
     2101 gcttctgacg ttcagtgcag ccgtcatctg aaaacgacat gtcgcacaag tcctaagtta
     2161 cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca
     2221 taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg
     2281 ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg
     2341 ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc
     2401 gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac gttgtgacag
     2461 tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt gccgagcgca
     2521 tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac accaccacgc
     2581 cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag cgttccctaa
     2641 tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc
     2701 cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc gaccaggaag
     2761 gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg
     2821 cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt gccttccgtg
     2881 aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc caagaggaac
     2941 aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg aagagatcga
     3001 ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacctct caaccgtgcg
     3061 gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt
     3121 ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag
     3181 cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg
     3241 gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc
     3301 gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc
     3361 gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt
     3421 gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc
     3481 gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc
     3541 gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg
     3601 gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc
     3661 gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg
     3721 tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc
     3781 gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag
     3841 gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga
     3901 gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa
     3961 cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg
     4021 aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc
     4081 tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga
     4141 attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg
     4201 gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgcctgc
     4261 cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac
     4321 catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg
     4381 ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc
     4441 aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt
     4501 cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg
     4561 acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc
     4621 gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt
     4681 ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt
     4741 cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg
     4801 tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc
     4861 agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc
     4921 gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta
     4981 gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag
     5041 ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc
     5101 ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg
     5161 ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg
     5221 ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc
     5281 cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc
     5341 gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc
     5401 aaattgccct agcaggggaa aaaggtcgaa aaggactctt tcctgtggat agcacgtaca
     5461 ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt
     5521 acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt
     5581 ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac
     5641 tgtctggcca gcgcacagcc gaagtgctgc aaaaagcgcc tacccttcgg tcgctgcgct
     5701 ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg
     5761 gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc
     5821 ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     5881 cacatgcagc tcccggtgac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5941 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     6001 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     6061 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     6121 ggcgctcatc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6721 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6841 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6901 atcctttgat c
//
