LOCUS       sequence-274961-        7303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acceptor plasmid for even level position 2, lacZ blue-white 
            screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7303)
  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 7303)
  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: pCsB
FEATURES             Location/Qualifiers
     source          1..7303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    134..158
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     499..521
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     513..530
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     514..530
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    531..587
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(600..616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(600..616)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(613..635)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    624..640
                     /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(648..654,655..672,673..678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    693..714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    798..822
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2123..2752
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3181..4254
                     /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      4320..4514
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4750..4772)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4858..4998
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5013..5030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5184..5772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5264..5283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5860..6651)
                     /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 gcatcggtaa catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc
       61 ggactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct
      121 gttggctggc tggtggcagg atatattgtg gtgtaaacat aacggatccg gtctcatact
      181 aatgtgaaga gccacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      241 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      301 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      361 ccgcatcagg cgctcatccg ccattcaggc tgcgcaactg ttgggaaggg cgatcggcgc
      421 aggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      481 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
      541 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
      601 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      661 ggaagcatat agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
      721 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctggc tcttcaggta
      781 aatgtgagac cggatcctga caggatatat tggcgggtaa acctaagaga aaagagcgtt
      841 tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta
      901 tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta ctttgatcca acccctccgc
      961 tgctatagtg cagtcggctt ctgacgttca gtgcagccgt catctgaaaa cgacatgtcg
     1021 cacaagtcct aagttacgcg acaggctgcc gccctgccct tttcctggcg ttttcttgtc
     1081 gcgtgtttta gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg
     1141 aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac
     1201 ttgaccaacc aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag
     1261 atcaccggca ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct
     1321 ggcgacgttg tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg
     1381 gacattgccg agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg
     1441 gccgacacca ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag
     1501 ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga
     1561 ggcgtgaagt ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag
     1621 ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc
     1681 tcgaccctgt accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg
     1741 cgcggtgcct tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat
     1801 gaacgccaag aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca
     1861 ttaccgaaga gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc
     1921 acctctcaac cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg
     1981 cctggccggc cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg
     2041 tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata
     2101 aacaaatacg caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc
     2161 aggcaagacg accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt
     2221 tctgttagtc gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga
     2281 tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat
     2341 cggccggcgc gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc
     2401 cgcgatcaag gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg
     2461 ggccaccgcc gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct
     2521 acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc
     2581 cgaggcgctg gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag
     2641 ctacccaggc actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc
     2701 tgcccgcgag gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga
     2761 ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc
     2821 agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac
     2881 tttcagttgc cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag
     2941 accattaccg agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga
     3001 ataaatgagt agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc
     3061 aggcaccgac gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag
     3121 cggctgggtt gcctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc
     3181 gtgagcggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg
     3241 gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc
     3301 cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg
     3361 gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc
     3421 gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt
     3481 ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac
     3541 gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg
     3601 gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga
     3661 gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga
     3721 gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg
     3781 cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag
     3841 ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac
     3901 atcgagatcg agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac
     3961 gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac
     4021 cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac
     4081 gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc
     4141 gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc
     4201 ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg
     4261 gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg actctttcct
     4321 gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt
     4381 gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag
     4441 aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc
     4501 ctggcctgtg cataactgtc tggccagcgc acagccgaag tgctgcaaaa agcgcctacc
     4561 cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc
     4621 cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg
     4681 tcgccactcg accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga
     4741 cggtgaaaac ctctgacaca tgcagctccc ggtgacggtc acagcttgtc tgtaagcgga
     4801 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     4861 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     4921 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     4981 agaaaatacc gcatcaggcg ctcatccgct tcctcgctca ctgactcgct gcgctcggtc
     5041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5161 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5581 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5821 aactcacgtt aagggatttt ggtcatgcat tctaggtgat tatttgccga ctaccttggt
     5881 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     5941 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     6001 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     6061 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcac cagcccagtc
     6121 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     6181 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     6241 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     6301 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     6361 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     6421 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     6481 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     6541 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     6601 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca taatgtttaa
     6661 ctttgtttta gggcgactgc cctgctgcgt aacatcgttg ctgctccata acatcaaaca
     6721 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtaccccaaa
     6781 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     6841 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagct tacgaaccga
     6901 acaggcttat gtccactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     6961 aaccttgggt agcagcgaag tcgaggcatt tctgtcctgg ctggaacaga acttattatt
     7021 tccttcctct tttctacagt atttaaagat accccaagaa gctaattata acaagacgaa
     7081 ctccaattca ctgttccttg cattctaaaa ccttaaatac cagaaaacag ctttttcaaa
     7141 gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga aaccgcggtg
     7201 atcacaggca gcaacgctct gtcatcgtta caatcaacat gctaccctcc gcgagatcat
     7261 ccgtgtttca aacccggcag cttagttgcc gttcttccga ata
//
