LOCUS       sequence-333755-        3076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acceptor plasmid for odd level position 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3076)
  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 3076)
  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: pCk2chlo
FEATURES             Location/Qualifiers
     source          1..3076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     711..733
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     725..742
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     726..742
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    743..799
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(812..828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(812..828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(825..847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    836..852
                     /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(860..866,867..884,885..890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    905..926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1152..1174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1260..1400
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1415..1432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1586..2173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1666..1685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2261..3076)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2261..3076)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2984..3003
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aacagaactt attatttcct tcctcttttc tacagtattt aaagataccc caagaagcta
       61 attataacaa gacgaactcc aattcactgt tccttgcatt ctaaaacctt aaataccaga
      121 aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa catagtatcg acggagccga
      181 ttttgaaacc gcggtgatca caggcagcaa cgctctgtca tcgttacaat caacatgcta
      241 ccctccgcga gatcatccgt gtttcaaacc cggcagctta gttgccgttc ttccgaatag
      301 catcggtaac atgagcaaag tctgccgcct tacaacggct ctcccgctga cgccgtcccg
      361 gactgatgaa ttcgcggccg cgctcttcag caaggagaga gacccacagc ttgtctgtaa
      421 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
      481 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
      541 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcatc cgccattcag
      601 gctgcgcaac tgttgggaag ggcgatcggc gcaggcctct tcgctattac gccagctggc
      661 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      721 acgttgtaaa acgacggcca gtgaattcga gctcggtacc cggggatcct ctagagtcga
      781 cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      841 atccgctcac aattccacac aacatacgag ccggaagcat atagtgtaaa gcctggggtg
      901 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      961 gaaacctgtc gtgccagctg gggtctctcg ctatactgaa gagcgcggcc gcctgcagtg
     1021 gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc
     1081 cgtcgccact cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat
     1141 gacggtgaaa acctctgaca catgcagctc ccggtgacgg tcacagcttg tctgtaagcg
     1201 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     1261 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     1321 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     1381 ggagaaaata ccgcatcagg cgctcatccg cttcctcgct cactgactcg ctgcgctcgg
     1441 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1501 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1561 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1621 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1681 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1741 tgtccgcctt tctccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     2041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2221 aaactcacgt taagggattt tggtcatgca ttctaggtga ttagaaaaac tcatcgagca
     2281 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     2341 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2401 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2461 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     2521 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     2581 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agtcgaaata
     2641 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     2701 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     2761 ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     2821 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     2881 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     2941 tcccatacaa tcggtagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     3001 acccatataa atcagcatcc atgttggaat ttaatcgcgg ccttgagcaa gacgtttccc
     3061 gttgaatatg gctcat
//
