LOCUS       sequence-274975-        6911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use when making odd level constructs (L3,...) if pCsC position 
            (position 3) 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: pCsC_spacer
FEATURES             Location/Qualifiers
     source          1..6911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(37..625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(117..136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(713..1504)
                     /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    2290..2314
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    2562..2586
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3887..4516
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4945..6018
                     /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      6084..6278
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6514..6536)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6622..6762
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6677..6696
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6777..6794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
       61 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      121 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      181 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      241 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      301 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      361 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      421 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
      481 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      541 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      601 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      661 tcagtggaac gaaaactcac gttaagggat tttggtcatg cattctaggt gattatttgc
      721 cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc
      781 gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg
      841 gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga
      901 ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat
      961 caccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata
     1021 gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc
     1081 tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga
     1141 agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg
     1201 gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa
     1261 tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg
     1321 ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca
     1381 ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc
     1441 gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc
     1501 tcataatgtt taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc
     1561 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     1621 actgtacccc aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     1681 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     1741 gcttacgaac cgaacaggct tatgtccact gggttcgtgc cttcatccgt ttccacggtg
     1801 tgcgtcaccc ggcaaccttg ggtagcagcg aagtcgaggc atttctgtcc tggctggaac
     1861 agaacttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
     1921 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa
     1981 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
     2041 tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc
     2101 tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat
     2161 cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac
     2221 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg
     2281 gctggctggt ggcaggatat attgtggtgt aaacataacg gatccggtct caaatgaatg
     2341 gtagacactt tattagttag atccgctgtt attctgcacg attgcgcttg ccggtggaaa
     2401 tttaaggacg agtgtagtgc actcaccctt agccttcagc gtgacaagat tagggtagca
     2461 ccggaaacag cgattttacg gacagatagt acagtctgag actcagtcac tggtccctaa
     2521 acgggagccg gtacgtcggc ggtagctttg agaccggatc ctgacaggat atattggcgg
     2581 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa
     2641 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc tcgggatcaa
     2701 agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag
     2761 ccgtcatctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg
     2821 cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac
     2881 tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc
     2941 cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc
     3001 tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc
     3061 aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg
     3121 gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc
     3181 ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg
     3241 accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc
     3301 gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg
     3361 gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg
     3421 gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa
     3481 gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc
     3541 gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg
     3601 acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg
     3661 ggtacgtgtt cgagccgccc gcgcacctct caaccgtgcg gctgcatgaa atcctggccg
     3721 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc
     3781 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
     3841 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
     3901 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
     3961 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
     4021 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
     4081 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
     4141 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
     4201 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
     4261 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
     4321 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
     4381 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
     4441 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
     4501 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
     4561 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
     4621 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
     4681 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
     4741 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
     4801 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
     4861 aacgggcggt tggccaggcg taagcggctg ggttgcctgc cggccctgca atggcactgg
     4921 aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc
     4981 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
     5041 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
     5101 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
     5161 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
     5221 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
     5281 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
     5341 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
     5401 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
     5461 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
     5521 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
     5581 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
     5641 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
     5701 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
     5761 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
     5821 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
     5881 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
     5941 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
     6001 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
     6061 aaaggtcgaa aaggactctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
     6121 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
     6181 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
     6241 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
     6301 gaagtgctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
     6361 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
     6421 ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc
     6481 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggtgac
     6541 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6601 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     6661 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     6721 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcatc cgcttcctcg
     6781 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6841 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6901 ggccagcaaa a
//
