LOCUS       sequence-275304-        9466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L1 in position 1 (next to LB in L2 construct), for CsR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9466)
  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 9466)
  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: L1_CsR-Ck1
FEATURES             Location/Qualifiers
     source          1..9466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    433..457
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        574..1250
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     1187..1207
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    3394..3568
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3589..3613
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4914..5543
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5972..7045
                     /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      7111..7305
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7541..7563)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7649..7789
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7704..7723
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7804..7821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7975..8563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8055..8074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8651..9466)
                     /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(8651..9466)
                     /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     9374..9393
                     /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 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
      421 ttggctggct ggtggcagga tatattgtgg tgtaaacata acgaattgct cttcaatgag
      481 gaggctagag cagcttgcca acatggtgga gcacgacact ctcgtctact ccaagaatat
      541 caaagataca gtctcagaag accaaagggc tattgagact tttcaacaaa gggtaatatc
      601 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa ggacagtaga
      661 aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga
      721 tgcctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
      781 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgaacatg gtggagcacg
      841 acactctcgt ctactccaag aatatcaaag atacagtctc agaagaccaa agggctattg
      901 agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct
      961 gtcacttcat caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc catcattgcg
     1021 ataaaggaaa ggctatcgtt caagatgcct ctgccgacag tggtcccaaa gatggacccc
     1081 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
     1141 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
     1201 acccttcctc tatataagga agttcatttc atttggagag gacacgctga aatcaccagt
     1261 ctctctctac aaatctatct ctctcgagct ttcgcagatc cggggggcaa tgagattcac
     1321 caatggcgtc aattagtgga tgcgcagctg tgcctttgag catagcgagc ctggagaagg
     1381 tgcagccatc ggcaaggtct tgttgcaagg gagctaaaga gctaaaggtg aggagtggtt
     1441 tcttctcggg agccgctctg aggggacatg catacaacac ctctcggaag actgagcgtt
     1501 cactggaagt gaaagcccaa gctgttagcg agaagatcca gtctgcagtc gatgtgagtg
     1561 gaccacggct tagcagcggg aagtcactcg ttgcaccaac cccgtccgac tttcagtctc
     1621 gctacaagga tgacgagccc cgcaaaggag ccgacatttt ggtggaggct cttgagcgag
     1681 aagggattag aacggtgttc gcttatcccg gcggagcgtc tatggagatt caccaggctc
     1741 ttacacgttc cagcagcatc cgcaacattc tctgtcgcca cgagcagggg gagattttcg
     1801 ctgctgaagg ttatgctaaa tcgtctggac gagtaggggt ttgtatagcc acatccggtc
     1861 ctggagcgac caatcttgtt accgggctcg ctgatgccct cttggacagt gtgccactag
     1921 tggccattac aggtcaagta tccagtagga tgattggtac cgatgctttt caagaaactc
     1981 caattgtgga ggttacgaga tcgatcacca agcacaatta ccttgttatg aacattgatg
     2041 acatccctcg cgtaatcagg gaggctttct ttttggcagc ttcaggtagg ccgggcccag
     2101 tcttggtaga tattcccaag gacgtgcaac agcaaatggc tattccagat tggtcgcagc
     2161 caatgcaact gaagggctac ttacaacgac tgcctcggcc accggaacca acgttaatcg
     2221 agcagattat aagactcatt tctagttcta agaagcctgt agtgtattct ggaggaggtt
     2281 gcaatcacgc cagcgaagag ttgagggaat ttgtcgagct taccggaatt cctatcacat
     2341 cgactttaat gggtttggga gcgtatcccg agtctgatcc attaagtttg aaaatgcttg
     2401 gaatgcatgg aactgtttac gccaactacg ccgtggacca agcagatctg ttgttggcat
     2461 ttggagtgcg tttcgacgat cgagtgactg gtaaactgga gacttttgct agcaggtcca
     2521 gcatcgttca tatcgatatt gaccctgctg aaattggaaa gaacaaacag ccccatgtgt
     2581 ccatgtgtgc tgatgtgaaa cctgcgctaa gtgaaatcaa tgccctcttg aagagtcaaa
     2641 agaggacttt cgacttctcc aactgggtgg cagaactaga agaacaaaag cagaaatggc
     2701 caatgaagtt tcccgtattc gaggatgcca ttgctcctca atctgccata cagacgctgt
     2761 acgaactcac tggtggaaat gccatcatat ctactggtgt tggccaacat caaatgtggg
     2821 cagcccagtg gttcaagtac aacgaaccgc ggcgttggtt gacatctgga ggtcttggag
     2881 ccatgggatt cggacttcct tctgctttgg gggctgccgc aactcaacca gatattcctg
     2941 tgatcgacat cgatggcgat ggtagctttg tgatgaacgt acaggagttg gctacaattt
     3001 ttgtggagaa gttacccgtg aagatcatga tcttaaacaa ccaacatcta ggaatggtcg
     3061 tacaattgga ggatagattc tacaaggcca acagagcaca tacttactta ggtgatccag
     3121 atcacgagga gcagatctac cccgacttct gtacagttgc agacggtttc aaggtgccaa
     3181 atgcgcgtgt ggtgaagaag gatgaactaa gagcagcaat tcaaactatg ctagattctc
     3241 ccggtcctta tttactggat gtgatagttc cccaccagga gcacgttctg cctatgattc
     3301 ctgggggagc aagttttaag gagattatca cagagggtga tggaaggact atatactaag
     3361 cttctctagc tagagtcgat cgacaagctc gagtttctcc ataataatgt gtgagtagtt
     3421 cccagataag ggaattaggg ttcctatagg gtttcgctca tgtgttgagc atataagaaa
     3481 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     3541 accaaaatcc agtactaaaa tccagatcgc tagcatgaag agcaattctg acaggatata
     3601 ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc
     3661 gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg
     3721 ggatcaaagt actttgatcc aacccctccg ctgctatagt gcagtcggct tctgacgttc
     3781 agtgcagccg tcatctgaaa acgacatgtc gcacaagtcc taagttacgc gacaggctgc
     3841 cgccctgccc ttttcctggc gttttcttgt cgcgtgtttt agtcgcataa agtagaatac
     3901 ttgcgactag aaccggagac attacgccat gaacaagagc gccgccgctg gcctgctggg
     3961 ctatgcccgc gtcagcaccg acgaccagga cttgaccaac caacgggccg aactgcacgc
     4021 ggccggctgc accaagctgt tttccgagaa gatcaccggc accaggcgcg accgcccgga
     4081 gctggccagg atgcttgacc acctacgccc tggcgacgtt gtgacagtga ccaggctaga
     4141 ccgcctggcc cgcagcaccc gcgacctact ggacattgcc gagcgcatcc aggaggccgg
     4201 cgcgggcctg cgtagcctgg cagagccgtg ggccgacacc accacgccgg ccggccgcat
     4261 ggtgttgacc gtgttcgccg gcattgccga gttcgagcgt tccctaatca tcgaccgcac
     4321 ccggagcggg cgcgaggccg ccaaggcccg aggcgtgaag tttggccccc gccctaccct
     4381 caccccggca cagatcgcgc acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa
     4441 agaggcggct gcactgcttg gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag
     4501 cgaggaagtg acgcccaccg aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac
     4561 cgaggccgac gccctggcgg ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg
     4621 caccaggacg gccaggacga accgtttttc attaccgaag agatcgaggc ggagatgatc
     4681 gcggccgggt acgtgttcga gccgcccgcg cacctctcaa ccgtgcggct gcatgaaatc
     4741 ctggccggtt tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa
     4801 accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg
     4861 gtcgctgcgt atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg
     4921 ttatcgctgt acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag
     4981 cccgcgccct gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca
     5041 gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc
     5101 gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg
     5161 gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga
     5221 ttccggtgca gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta
     5281 agcagcgcat tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga
     5341 tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca
     5401 ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa
     5461 ccgttcttga atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg
     5521 aaattaaatc aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa
     5581 cacgctaagt gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct
     5641 ggcagacacg ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa
     5701 gctgaagatg tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat
     5761 cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta
     5821 aaggaggcgg catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt
     5881 gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt tgcctgccgg ccctgcaatg
     5941 gcactggaac ccccaagccc gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt
     6001 acaaatcggc gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc
     6061 ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga
     6121 tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt gcgccgtcga ttaggaagcc
     6181 gcccaagggc gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg
     6241 cgatagtcgc agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc
     6301 tggcgaggtg atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc
     6361 cggcatggcc agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga
     6421 atccatgaac cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca
     6481 cgttgcggac gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct
     6541 ggtagaaacc tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc
     6601 caagaacggc cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat
     6661 cgtaaagagc gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta
     6721 ccgcgagatc acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt
     6781 gatcgatccc ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc
     6841 agaagccaga tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa
     6901 gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt
     6961 gaaggaggag gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga
     7021 gggcgaagca tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc
     7081 aggggaaaaa ggtcgaaaag gactctttcc tgtggatagc acgtacattg ggaacccaaa
     7141 gccgtacatt gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg
     7201 gtcacacatg taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc
     7261 tttaaaactt attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg
     7321 cacagccgaa gtgctgcaaa aagcgcctac ccttcggtcg ctgcgctccc tacgccccgc
     7381 cgcttcgcgt cggcctatcg cggccgctgg ccgctcaaaa atggctggcc tacggccagg
     7441 caatctacca gggcgcggac aagccgcgcc gtcgccactc gaccgccggc gcccacatca
     7501 aggcaccctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     7561 cggtgacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     7621 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     7681 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     7741 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcatccgc
     7801 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7861 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7921 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7981 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     8041 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     8101 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     8161 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     8221 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8281 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8341 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8401 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8461 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8521 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8581 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgca
     8641 ttctaggtga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     8701 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     8761 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     8821 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     8881 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     8941 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     9001 ttcgtgattg cgcctgagcg agtcgaaata cgcgatcgct gttaaaagga caattacaaa
     9061 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     9121 aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca gtggtgagta
     9181 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     9241 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     9301 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcggtagatt gtcgcacctg
     9361 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     9421 ttaatcgcgg ccttgagcaa gacgtttccc gttgaatatg gctcat
//
