LOCUS       sequence-274960-        7303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acceptor plasmid for even level position 1, 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: pCsA
FEATURES             Location/Qualifiers
     source          1..7303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    73..97
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     438..460
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     452..469
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     453..469
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    470..526
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(539..555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(539..555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(552..574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    563..579
                     /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(587..593,594..611,612..617))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    632..653
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    737..761
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2062..2691
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3120..4193
                     /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      4259..4453
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4689..4711)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4797..4937
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4952..4969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5123..5711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5203..5222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5799..6590)
                     /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 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
       61 ttggctggct ggtggcagga tatattgtgg tgtaaacata acggatccgg tctcaggaga
      121 atgtgaagag ccacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      181 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      241 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      301 cgcatcaggc gctcatccgc cattcaggct gcgcaactgt tgggaagggc gatcggcgca
      361 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      421 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
      481 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt
      541 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      601 gaagcatata gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
      661 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctggct cttcaggtat
      721 acttgagacc ggatcctgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt
      781 attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat
      841 gtgcatgcca accacagggt tcccctcggg atcaaagtac tttgatccaa cccctccgct
      901 gctatagtgc agtcggcttc tgacgttcag tgcagccgtc atctgaaaac gacatgtcgc
      961 acaagtccta agttacgcga caggctgccg ccctgccctt ttcctggcgt tttcttgtcg
     1021 cgtgttttag tcgcataaag tagaatactt gcgactagaa ccggagacat tacgccatga
     1081 acaagagcgc cgccgctggc ctgctgggct atgcccgcgt cagcaccgac gaccaggact
     1141 tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac caagctgttt tccgagaaga
     1201 tcaccggcac caggcgcgac cgcccggagc tggccaggat gcttgaccac ctacgccctg
     1261 gcgacgttgt gacagtgacc aggctagacc gcctggcccg cagcacccgc gacctactgg
     1321 acattgccga gcgcatccag gaggccggcg cgggcctgcg tagcctggca gagccgtggg
     1381 ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc attgccgagt
     1441 tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc aaggcccgag
     1501 gcgtgaagtt tggcccccgc cctaccctca ccccggcaca gatcgcgcac gcccgcgagc
     1561 tgatcgacca ggaaggccgc accgtgaaag aggcggctgc actgcttggc gtgcatcgct
     1621 cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag gccaggcggc
     1681 gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc cctggcggcc gccgagaatg
     1741 aacgccaaga ggaacaagca tgaaaccgca ccaggacggc caggacgaac cgtttttcat
     1801 taccgaagag atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc cgcccgcgca
     1861 cctctcaacc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca agctggcggc
     1921 ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt
     1981 atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa
     2041 acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa aggcgggtca
     2101 ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt
     2161 ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt gcgggaagat
     2221 caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt gaaggccatc
     2281 ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt ggctgtgtcc
     2341 gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta cgacatatgg
     2401 gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga tggaaggcta
     2461 caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc
     2521 gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc
     2581 tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga gggcgacgct
     2641 gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg agttaatgag
     2701 gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg agcgcacgca
     2761 gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag cgggtcaact
     2821 ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc caaggcaaga
     2881 ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg agcaaatgaa
     2941 taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca agaacaacca
     3001 ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc
     3061 ggctgggttg cctgccggcc ctgcaatggc actggaaccc ccaagcccga ggaatcggcg
     3121 tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt gatgacctgg
     3181 tggagaagtt gaaggccgcg caggccgccc agcggcaacg catcgaggca gaagcacgcc
     3241 ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg caaccgccgg
     3301 cagccggtgc gccgtcgatt aggaagccgc ccaagggcga cgagcaacca gattttttcg
     3361 ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac gtggccgttt
     3421 tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag cttccagacg
     3481 ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat tacgacctgg
     3541 tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa gggaagggag
     3601 acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc tgccggcgag
     3661 ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta aacaccacgc
     3721 acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg gtatccgagg
     3781 gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg ccggagtaca
     3841 tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag aacccggacg
     3901 tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt tttctctacc
     3961 gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag acgatctacg
     4021 aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc aagctgatcg
     4081 ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct ggcccgatcc
     4141 tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg
     4201 agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaagga ctctttcctg
     4261 tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac ccgtacattg
     4321 ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat ataaaagaga
     4381 aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt aaaacccgcc
     4441 tggcctgtgc ataactgtct ggccagcgca cagccgaagt gctgcaaaaa gcgcctaccc
     4501 ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg gccgctggcc
     4561 gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa gccgcgccgt
     4621 cgccactcga ccgccggcgc ccacatcaag gcaccctgcc tcgcgcgttt cggtgatgac
     4681 ggtgaaaacc tctgacacat gcagctcccg gtgacggtca cagcttgtct gtaagcggat
     4741 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     4801 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     4861 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4921 gaaaataccg catcaggcgc tcatccgctt cctcgctcac tgactcgctg cgctcggtcg
     4981 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5041 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5101 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5161 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5221 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5281 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5341 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5401 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5461 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5521 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5581 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5641 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5701 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5761 actcacgtta agggattttg gtcatgcatt ctaggtgatt atttgccgac taccttggtg
     5821 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
     5881 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
     5941 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
     6001 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcacc agcccagtcg
     6061 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
     6121 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
     6181 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
     6241 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
     6301 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
     6361 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
     6421 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact
     6481 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
     6541 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat aatgtttaac
     6601 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctccataa catcaaacat
     6661 cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg taccccaaaa
     6721 aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc
     6781 aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagctt acgaaccgaa
     6841 caggcttatg tccactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca
     6901 accttgggta gcagcgaagt cgaggcattt ctgtcctggc tggaacagaa cttattattt
     6961 ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac
     7021 tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc tttttcaaag
     7081 ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa accgcggtga
     7141 tcacaggcag caacgctctg tcatcgttac aatcaacatg ctaccctccg cgagatcatc
     7201 cgtgtttcaa acccggcagc ttagttgccg ttcttccgaa tagcatcggt aacatgagca
     7261 aagtctgccg ccttacaacg gctctcccgc tgacgccgtc ccg
//
