LOCUS       sequence-274976-        6911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use when making odd level constructs (L3,...) if pCsE position 
            (position 4) 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: pCsE_spacer
FEATURES             Location/Qualifiers
     source          1..6911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(203..994)
                     /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    1780..1804
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    2052..2076
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3377..4006
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4435..5508
                     /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      5574..5768
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6004..6026)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6112..6252
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6167..6186
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6267..6284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6438..6911,1..115))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6518..6537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
       61 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      181 tttggtcatg cattctaggt gattatttgc cgactacctt ggtgatctcg cctttcacgt
      241 agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa
      301 gataagcctg tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg
      361 cccagtcggc agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc
      421 gggacaacgt aagcactaca tttcgctcat caccagccca gtcgggcggc gagttccata
      481 gcgttaaggt ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt
      541 cctccgccgc tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag
      601 ccagatcaat gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc
      661 attctccaaa ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca
      721 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
      781 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
      841 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
      901 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
      961 gagtcgatac ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac
     1021 tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
     1081 gcgcttgctg cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag
     1141 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
     1201 tgcgtgagcg catacgctac ttgcattaca gcttacgaac cgaacaggct tatgtccact
     1261 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggtagcagcg
     1321 aagtcgaggc atttctgtcc tggctggaac agaacttatt atttccttcc tcttttctac
     1381 agtatttaaa gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc
     1441 ttgcattcta aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg
     1501 cgtataacat agtatcgacg gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc
     1561 tctgtcatcg ttacaatcaa catgctaccc tccgcgagat catccgtgtt tcaaacccgg
     1621 cagcttagtt gccgttcttc cgaatagcat cggtaacatg agcaaagtct gccgccttac
     1681 aacggctctc ccgctgacgc cgtcccggac tgatgggctg cctgtatcga gtggtgattt
     1741 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt
     1801 aaacataacg gatccggtct cagcttaatg gtagacactt tattagttag atccgctgtt
     1861 attctgcacg attgcgcttg ccggtggaaa tttaaggacg agtgtagtgc actcaccctt
     1921 agccttcagc gtgacaagat tagggtagca ccggaaacag cgattttacg gacagatagt
     1981 acagtctgag actcagtcac tggtccctaa acgggagccg gtacgtcggc ggttcgcttg
     2041 agaccggatc ctgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag
     2101 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca
     2161 tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat
     2221 agtgcagtcg gcttctgacg ttcagtgcag ccgtcatctg aaaacgacat gtcgcacaag
     2281 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
     2341 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
     2401 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
     2461 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
     2521 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
     2581 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
     2641 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
     2701 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
     2761 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
     2821 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
     2881 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
     2941 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
     3001 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
     3061 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
     3121 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacctct
     3181 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
     3241 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     3301 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
     3361 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     3421 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     3481 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     3541 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     3601 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     3661 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     3721 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     3781 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     3841 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     3901 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     3961 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     4021 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     4081 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     4141 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     4201 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     4261 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     4321 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     4381 ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     4441 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     4501 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     4561 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     4621 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     4681 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     4741 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     4801 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     4861 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     4921 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     4981 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     5041 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     5101 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     5161 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     5221 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     5281 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     5341 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     5401 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     5461 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     5521 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggactctt tcctgtggat
     5581 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     5641 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     5701 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     5761 tgtgcataac tgtctggcca gcgcacagcc gaagtgctgc aaaaagcgcc tacccttcgg
     5821 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     5881 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     5941 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     6001 aaacctctga cacatgcagc tcccggtgac ggtcacagct tgtctgtaag cggatgccgg
     6061 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     6121 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     6181 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     6241 taccgcatca ggcgctcatc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6841 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6901 ctgctgaagc c
//
