LOCUS       sequence-274962-        7303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acceptor plasmid for even level position 3, 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: pCsC
FEATURES             Location/Qualifiers
     source          1..7303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    134..158
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    242..263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(278..283,284..301,302..308)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    316..332
                     /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)."
     primer_bind     321..343
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     340..356
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     340..356
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    369..425
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(426..443)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(426..442)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(435..457)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    798..822
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             1608..2399
                     /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"
     rep_origin      2487..3075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2976..2995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3229..3246
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3261..3401
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3487..3509
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      3745..3939
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(4005..5078)
                     /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"
     CDS             complement(5507..6136)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 ggggttggat caaagtactt tgatcccgag gggaaccctg tggttggcat gcacatacaa
       61 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
      121 tcttttctct taggtttacc cgccaatata tcctgtcagg atccggtctc aaagctacct
      181 gaagagccag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
      241 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactatatg cttccggctc
      301 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
      361 attacgccaa gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg
      421 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      481 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcctgcgcc
      541 gatcgccctt cccaacagtt gcgcagcctg aatggcggat gagcgcctga tgcggtattt
      601 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
      661 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
      721 acgggcttgt ctgctcccgg catccgctta cagacaagct gtggctcttc acattcattt
      781 gagaccggat ccgttatgtt tacaccacaa tatatcctgc caccagccag ccaacagctc
      841 cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc agtccgggac
      901 ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc gatgctattc
      961 ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg agggtagcat
     1021 gttgattgta acgatgacag agcgttgctg cctgtgatca ccgcggtttc aaaatcggct
     1081 ccgtcgatac tatgttatac gccaactttg aaaacaactt tgaaaaagct gttttctggt
     1141 atttaaggtt ttagaatgca aggaacagtg aattggagtt cgtcttgtta taattagctt
     1201 cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataagttc tgttccagcc
     1261 aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga
     1321 aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca
     1381 agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg
     1441 gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc
     1501 gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca
     1561 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg
     1621 tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg
     1681 aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac
     1741 acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag
     1801 ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg
     1861 ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc
     1921 gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag
     1981 ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct
     2041 tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg
     2101 cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggt gatgagcgaa
     2161 atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga
     2221 aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac
     2281 ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc
     2341 agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataat
     2401 cacctagaat gcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2461 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2521 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2581 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2641 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2701 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2761 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2821 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2881 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2941 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3001 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3061 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3121 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3181 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3241 agcgaggaag cggatgagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3301 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3361 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3421 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3481 ccgtcaccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3541 agggtgcctt gatgtgggcg ccggcggtcg agtggcgacg gcgcggcttg tccgcgccct
     3601 ggtagattgc ctggccgtag gccagccatt tttgagcggc cagcggccgc gataggccga
     3661 cgcgaagcgg cggggcgtag ggagcgcagc gaccgaaggg taggcgcttt ttgcagcact
     3721 tcggctgtgc gctggccaga cagttatgca caggccaggc gggttttaag agttttaata
     3781 agttttaaag agttttaggc ggaaaaatcg ccttttttct cttttatatc agtcacttac
     3841 atgtgtgacc ggttcccaat gtacggcttt gggttcccaa tgtacgggtt ccggttccca
     3901 atgtacggct ttgggttccc aatgtacgtg ctatccacag gaaagagtcc ttttcgacct
     3961 ttttcccctg ctagggcaat ttgccctagc atctgctccg tacattagga accggcggat
     4021 gcttcgccct cgatcaggtt gcggtagcgc atgactagga tcgggccagc ctgccccgcc
     4081 tcctccttca aatcgtactc cggcaggtca tttgacccga tcagcttgcg cacggtgaaa
     4141 cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt agatcgtctt gaacaaccat
     4201 ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt agagaaaacg gccgatgccg
     4261 ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt ccgggttctt gccttctgtg
     4321 atctcgcggt acatccaatc agctagctcg atctcgatgt actccggccg cccggtttcg
     4381 ctctttacga tcttgtagcg gctaatcaag gcttcaccct cggataccgt caccaggcgg
     4441 ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg tggtgtttaa ccgaatgcag
     4501 gtttctacca ggtcgtcttt ctgctttccg ccatcggctc gccggcagaa cttgagtacg
     4561 tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc ccttcccttc ccggtatcgg
     4621 ttcatggatt cggttagatg ggaaaccgcc atcagtacca ggtcgtaatc ccacacactg
     4681 gccatgccgg ccggccctgc ggaaacctct acgtgcccgt ctggaagctc gtagcggatc
     4741 acctcgccag ctcgtcggtc acgcttcgac agacggaaaa cggccacgtc catgatgctg
     4801 cgactatcgc gggtgcccac gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg
     4861 cccttgggcg gcttcctaat cgacggcgca ccggctgccg gcggttgccg ggattctttg
     4921 cggattcgat cagcggccgc ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt
     4981 tgccgctggg cggcctgcgc ggccttcaac ttctccacca ggtcatcacc cagcgccgcg
     5041 ccgatttgta ccgggccgga tggtttgcga ccgctcacgc cgattcctcg ggcttggggg
     5101 ttccagtgcc attgcagggc cggcaggcaa cccagccgct tacgcctggc caaccgcccg
     5161 ttcctccaca catggggcat tccacggcgt cggtgcctgg ttgttcttga ttttccatgc
     5221 cgcctccttt agccgctaaa attcatctac tcatttattc atttgctcat ttactctggt
     5281 agctgcgcga tgtattcaga tagcagctcg gtaatggtct tgccttggcg taccgcgtac
     5341 atcttcagct tggtgtgatc ctccgccggc aactgaaagt tgacccgctt catggctggc
     5401 gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc gtgcgctcgg acggccggca
     5461 cttagcgtgt ttgtgctttt gctcattttc tctttacctc attaactcaa atgagttttg
     5521 atttaatttc agcggccagc gcctggacct cgcgggcagc gtcgccctcg ggttctgatt
     5581 caagaacggt tgtgccggcg gcggcagtgc ctgggtagct cacgcgctgc gtgatacggg
     5641 actcaagaat gggcagctcg tacccggcca gcgcctcggc aacctcaccg ccgatgcgcg
     5701 tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag ccttccatcc gtgacctcaa
     5761 tgcgctgctt aaccagctcc accaggtcgg cggtggccca tatgtcgtaa gggcttggct
     5821 gcaccggaat cagcacgaag tcggctgcct tgatcgcgga cacagccaag tccgccgcct
     5881 ggggcgctcc gtcgatcact acgaagtcgc gccggccgat ggccttcacg tcgcggtcaa
     5941 tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc ttcccgcacg gccgcccaat
     6001 cgcgggcact gccctgggga tcggaatcga ctaacagaac atcggccccg gcgagttgca
     6061 gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga cccgcctttc tggttaagta
     6121 cagcgataac cttcatgcgt tccccttgcg tatttgttta tttactcatc gcatcatata
     6181 cgcagcgacc gcatgacgca agctgtttta ctcaaataca catcaccttt ttagacggcg
     6241 gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc gccagcttgg catcagacaa
     6301 accggccagg atttcatgca gccgcacggt tgagaggtgc gcgggcggct cgaacacgta
     6361 cccggccgcg atcatctccg cctcgatctc ttcggtaatg aaaaacggtt cgtcctggcc
     6421 gtcctggtgc ggtttcatgc ttgttcctct tggcgttcat tctcggcggc cgccagggcg
     6481 tcggcctcgg tcaatgcgtc ctcacggaag gcaccgcgcc gcctggcctc ggtgggcgtc
     6541 acttcctcgc tgcgctcaag tgcgcggtac agggtcgagc gatgcacgcc aagcagtgca
     6601 gccgcctctt tcacggtgcg gccttcctgg tcgatcagct cgcgggcgtg cgcgatctgt
     6661 gccggggtga gggtagggcg ggggccaaac ttcacgcctc gggccttggc ggcctcgcgc
     6721 ccgctccggg tgcggtcgat gattagggaa cgctcgaact cggcaatgcc ggcgaacacg
     6781 gtcaacacca tgcggccggc cggcgtggtg gtgtcggccc acggctctgc caggctacgc
     6841 aggcccgcgc cggcctcctg gatgcgctcg gcaatgtcca gtaggtcgcg ggtgctgcgg
     6901 gccaggcggt ctagcctggt cactgtcaca acgtcgccag ggcgtaggtg gtcaagcatc
     6961 ctggccagct ccgggcggtc gcgcctggtg ccggtgatct tctcggaaaa cagcttggtg
     7021 cagccggccg cgtgcagttc ggcccgttgg ttggtcaagt cctggtcgtc ggtgctgacg
     7081 cgggcatagc ccagcaggcc agcggcggcg ctcttgttca tggcgtaatg tctccggttc
     7141 tagtcgcaag tattctactt tatgcgacta aaacacgcga caagaaaacg ccaggaaaag
     7201 ggcagggcgg cagcctgtcg cgtaacttag gacttgtgcg acatgtcgtt ttcagatgac
     7261 ggctgcactg aacgtcagaa gccgactgca ctatagcagc gga
//
