LOCUS       sequence-332028-        3404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acceptor plasmid for even level position 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3404)
  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 3404)
  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: pCsBchlo
FEATURES             Location/Qualifiers
     source          1..3404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      190..777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     678..697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     931..948
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    963..1103
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1189..1211
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1688..1710
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1702..1719
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1703..1719
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1720..1776
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1789..1805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1789..1805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1802..1824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1813..1829
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1837..1843,1844..1861,1862..1867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1882..1903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             join(2715..3404,1..102)
                     /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 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
       61 cactacgtga aaggcgagat caccaaggta gtcggcaaat aatcacctag aatgcatgac
      121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      301 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
      361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      601 tcccgaaggg agaaaggcgg acaggtatcg gtaagcggca gggtcggaac aggagagcgc
      661 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      721 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      781 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      841 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      901 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggatgag
      961 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     1021 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     1081 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     1141 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtcacc gggagctgca
     1201 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagggtgcc ttgatgtggg
     1261 cgccggcggt cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt gcctggccgt
     1321 aggccagcca tttttgagcg gccactgcag gcggccgcgg tctcatacta atgtgaagag
     1381 ccacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     1441 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     1501 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     1561 gctcatccgc cattcaggct gcgcaactgt tgggaagggc gatcggcgca ggcctcttcg
     1621 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1681 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
     1741 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     1801 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcatata
     1861 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1921 gcccgctttc cagtcgggaa acctgtcgtg ccagctggct cttcaggtaa atgtgagacc
     1981 gcggccgcga attcatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag
     2041 actttgctca tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa
     2101 cacggatgat ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct
     2161 gtgatcaccg cggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa
     2221 acaactttga aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat
     2281 tggagttcgt cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg
     2341 aaggaaataa taagttctgt tccagccagg acagaaatgc ctcgacttcg ctgctaccca
     2401 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     2461 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     2521 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     2581 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     2641 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     2701 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     2761 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     2821 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     2881 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     2941 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     3001 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     3061 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     3121 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     3181 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     3241 cgcccgactg ggctggtgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     3301 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     3361 tgccggccca gtatcagccc gtcatacttg aagctagaca ggct
//
