LOCUS       sequence-274963-        7303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acceptor plasmid for even level position 4, 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: pCsE
FEATURES             Location/Qualifiers
     source          1..7303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1136..1765
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2194..3267
                     /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      3333..3527
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3763..3785)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3871..4011
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4026..4043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4197..4785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4277..4296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4873..5664)
                     /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    6450..6474
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     6815..6837
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6829..6846
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6830..6846
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6847..6903
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(6916..6932)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6916..6932)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6929..6951)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6940..6956
                     /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(6964..6970,6971..6988,6989..6994))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7009..7030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7114..7138
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tcagtgcagc cgtcatctga aaacgacatg tcgcacaagt cctaagttac gcgacaggct
       61 gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat
      121 acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc tggcctgctg
      181 ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc cgaactgcac
      241 gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg cgaccgcccg
      301 gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt gaccaggcta
      361 gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc
      421 ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc
      481 atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat catcgaccgc
      541 acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc ccgccctacc
      601 ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg
      661 aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc
      721 agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga ggacgcattg
      781 accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca agcatgaaac
      841 cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag gcggagatga
      901 tcgcggccgg gtacgtgttc gagccgcccg cgcacctctc aaccgtgcgg ctgcatgaaa
      961 tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg gccgctgaag
     1021 aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg
     1081 cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa
     1141 ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg caacccatct
     1201 agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg atccccaggg
     1261 cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga
     1321 ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg tagtgatcga
     1381 cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg acttcgtgct
     1441 gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg tggagctggt
     1501 taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc
     1561 gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc
     1621 cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg ccgccggcac
     1681 aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg cgctggccgc
     1741 tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag caaaagcaca
     1801 aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac gttggccagc
     1861 ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga ggatcacacc
     1921 aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct atctgaatac
     1981 atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa ttttagcggc
     2041 taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg aatgccccat
     2101 gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc ggccctgcaa
     2161 tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc atccggcccg
     2221 gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc cgcgcaggcc
     2281 gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca agcggccgct
     2341 gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc gattaggaag
     2401 ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga cgtgggcacc
     2461 cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg tgaccgacga
     2521 gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc cgcagggccg
     2581 gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc ccatctaacc
     2641 gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt gttccgtcca
     2701 cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca gaaagacgac
     2761 ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg tacgaagaag
     2821 gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag ccgctacaag
     2881 atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc tgattggatg
     2941 taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc cgattacttt
     3001 ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc cgcaggcaag
     3061 gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc cggagagttc
     3121 aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc ggagtacgat
     3181 ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg caacctgatc
     3241 gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca aattgcccta
     3301 gcaggggaaa aaggtcgaaa aggactcttt cctgtggata gcacgtacat tgggaaccca
     3361 aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta cattgggaac
     3421 cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc cgcctaaaac
     3481 tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact gtctggccag
     3541 cgcacagccg aagtgctgca aaaagcgcct acccttcggt cgctgcgctc cctacgcccc
     3601 gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg cctacggcca
     3661 ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg gcgcccacat
     3721 caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3781 cccggtgacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3841 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     3901 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     3961 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcatcc
     4021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4621 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4861 cattctaggt gattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     4921 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     4981 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     5041 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     5101 aagcactaca tttcgctcat caccagccca gtcgggcggc gagttccata gcgttaaggt
     5161 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     5221 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     5281 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     5341 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     5401 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     5461 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     5521 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     5581 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     5641 ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac tgccctgctg
     5701 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     5761 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac
     5821 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     5881 catacgctac ttgcattaca gcttacgaac cgaacaggct tatgtccact gggttcgtgc
     5941 cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggtagcagcg aagtcgaggc
     6001 atttctgtcc tggctggaac agaacttatt atttccttcc tcttttctac agtatttaaa
     6061 gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc ttgcattcta
     6121 aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg cgtataacat
     6181 agtatcgacg gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg
     6241 ttacaatcaa catgctaccc tccgcgagat catccgtgtt tcaaacccgg cagcttagtt
     6301 gccgttcttc cgaatagcat cggtaacatg agcaaagtct gccgccttac aacggctctc
     6361 ccgctgacgc cgtcccggac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc
     6421 tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacataacg
     6481 gatccggtct cagcttaatg tgaagagcca cagcttgtct gtaagcggat gccgggagca
     6541 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     6601 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     6661 gcgtaaggag aaaataccgc atcaggcgct catccgccat tcaggctgcg caactgttgg
     6721 gaagggcgat cggcgcaggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     6781 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     6841 gccagtgaat tcgagctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag
     6901 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6961 acacaacata cgagccggaa gcatatagtg taaagcctgg ggtgcctaat gagtgagcta
     7021 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7081 gctggctctt caggttcgct tgagaccgga tcctgacagg atatattggc gggtaaacct
     7141 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
     7201 cgttcgtcca tttgtatgtg catgccaacc acagggttcc cctcgggatc aaagtacttt
     7261 gatccaaccc ctccgctgct atagtgcagt cggcttctga cgt
//
