LOCUS       sequence-275251-        8479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  L1 in position 1 (next to LB in L2 construct), for HygR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8479)
  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 8479)
  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_HyR-Ck1
FEATURES             Location/Qualifiers
     source          1..8479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(450..1265)
                     /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(450..1265)
                     /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     1173..1192
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    1698..1722
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2236..2581
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2524..2544
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2612..3637
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    3672..3846
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3867..3891
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5192..5821
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6250..7323
                     /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      7389..7583
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7819..7841)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7927..8067
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7982..8001
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8082..8099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8253..8479,1..362))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8333..8352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
       61 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      181 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
      241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      421 aagggatttt ggtcatgcat tctaggtgat tagaaaaact catcgagcat caaatgaaac
      481 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
      541 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
      601 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
      661 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
      721 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
      781 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gtcgaaatac gcgatcgctg
      841 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
      901 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct
      961 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     1021 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     1081 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     1141 cggtagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     1201 tcagcatcca tgttggaatt taatcgcggc cttgagcaag acgtttcccg ttgaatatgg
     1261 ctcataacag aacttattat ttccttcctc ttttctacag tatttaaaga taccccaaga
     1321 agctaattat aacaagacga actccaattc actgttcctt gcattctaaa accttaaata
     1381 ccagaaaaca gctttttcaa agttgttttc aaagttggcg tataacatag tatcgacgga
     1441 gccgattttg aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca
     1501 tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc cgttcttccg
     1561 aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc gctgacgccg
     1621 tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg
     1681 agctgttggc tggctggtgg caggatatat tgtggtgtaa acataacgaa ttgctcttca
     1741 atgaggagag attagccttt tcaatttcag aaagaatgct aacccacaga tggttagaga
     1801 ggcttacgca gcaagtctca tcaagacgat ctacccgagc aataatctac aggaaatcaa
     1861 ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa
     1921 cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg
     1981 cttgcttcac aaaccaaggc aagtaataga gattggagtc tctaaaaagg tagttcccac
     2041 tgaatcaaag gccatggagt caaagattca aatagaggac ctaacagaac tcgccgtaaa
     2101 gactggcgaa cagttcatac agagtctctt acgactcaat gacaagaaga aaatcttcgt
     2161 caacatggtg gagcacgaca cacttgtcta ctccaaaaat atcaaagata cagtctcagt
     2221 agaccaaagg gcaattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt
     2281 ccattgccca gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta
     2341 caaatgccat cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg
     2401 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaaaacg ttccaaccac
     2461 gtctacaaag caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc
     2521 ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagagaac
     2581 acgggggact ctagaggatc caacatcacc aatgaaaaag cctgaactca ccgcgacgtc
     2641 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     2701 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     2761 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
     2821 cgcgctcccg attccggaag tgcttgacat tggggagttt agcgagagcc tgacctattg
     2881 catctcccgc cgttcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     2941 tgttctacaa ccggtcgcgg aggctatgga tgcgatcgct gcggccgatc ttagccagac
     3001 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3061 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3121 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     3181 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     3241 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     3301 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     3361 gcggaggcat ccggagcttg caggatcgcc acgactccgg gcgtatatgc tccgcattgg
     3421 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     3481 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     3541 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     3601 ccgacgcccc agcactcgtc cgagggcaaa gaaataggct tctctagcta gagtcgatcg
     3661 acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg aattagggtt
     3721 cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt
     3781 gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc
     3841 cagatcgcta gcatgaagag caattctgac aggatatatt ggcgggtaaa cctaagagaa
     3901 aagagcgttt attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt
     3961 ccatttgtat gtgcatgcca accacagggt tcccctcggg atcaaagtac tttgatccaa
     4021 cccctccgct gctatagtgc agtcggcttc tgacgttcag tgcagccgtc atctgaaaac
     4081 gacatgtcgc acaagtccta agttacgcga caggctgccg ccctgccctt ttcctggcgt
     4141 tttcttgtcg cgtgttttag tcgcataaag tagaatactt gcgactagaa ccggagacat
     4201 tacgccatga acaagagcgc cgccgctggc ctgctgggct atgcccgcgt cagcaccgac
     4261 gaccaggact tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac caagctgttt
     4321 tccgagaaga tcaccggcac caggcgcgac cgcccggagc tggccaggat gcttgaccac
     4381 ctacgccctg gcgacgttgt gacagtgacc aggctagacc gcctggcccg cagcacccgc
     4441 gacctactgg acattgccga gcgcatccag gaggccggcg cgggcctgcg tagcctggca
     4501 gagccgtggg ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt gttcgccggc
     4561 attgccgagt tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg cgaggccgcc
     4621 aaggcccgag gcgtgaagtt tggcccccgc cctaccctca ccccggcaca gatcgcgcac
     4681 gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag aggcggctgc actgcttggc
     4741 gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac gcccaccgag
     4801 gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc cctggcggcc
     4861 gccgagaatg aacgccaaga ggaacaagca tgaaaccgca ccaggacggc caggacgaac
     4921 cgtttttcat taccgaagag atcgaggcgg agatgatcgc ggccgggtac gtgttcgagc
     4981 cgcccgcgca cctctcaacc gtgcggctgc atgaaatcct ggccggtttg tctgatgcca
     5041 agctggcggc ctggccggcc agcttggccg ctgaagaaac cgagcgccgc cgtctaaaaa
     5101 ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat atgatgcgat
     5161 gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac ttaaccagaa
     5221 aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc aactcgccgg
     5281 ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt gggcggccgt
     5341 gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg accgcgacgt
     5401 gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg cggcggactt
     5461 ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc caagccctta
     5521 cgacatatgg gccaccgccg acctggtgga gctggttaag cagcgcattg aggtcacgga
     5581 tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc gcatcggcgg
     5641 tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt cttgagtccc gtatcacgca
     5701 gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc gttcttgaat cagaacccga
     5761 gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa aactcatttg
     5821 agttaatgag gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc cggccgtccg
     5881 agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc agccatgaag
     5941 cgggtcaact ttcagttgcc ggcggaggat cacaccaagc tgaagatgta cgcggtacgc
     6001 caaggcaaga ccattaccga gctgctatct gaatacatcg cgcagctacc agagtaaatg
     6061 agcaaatgaa taaatgagta gatgaatttt agcggctaaa ggaggcggca tggaaaatca
     6121 agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg gcggttggcc
     6181 aggcgtaagc ggctgggttg cctgccggcc ctgcaatggc actggaaccc ccaagcccga
     6241 ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc ggcgctgggt
     6301 gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg catcgaggca
     6361 gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa agaatcccgg
     6421 caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga cgagcaacca
     6481 gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac
     6541 gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag
     6601 cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat
     6661 tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa
     6721 gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc
     6781 tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta
     6841 aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg
     6901 gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg
     6961 ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag
     7021 aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt
     7081 tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag
     7141 acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc
     7201 aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct
     7261 ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc
     7321 taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaagga
     7381 ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac
     7441 ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat
     7501 ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt
     7561 aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaagt gctgcaaaaa
     7621 gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg
     7681 gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa
     7741 gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc tcgcgcgttt
     7801 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gtgacggtca cagcttgtct
     7861 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     7921 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     7981 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     8041 tgcgtaagga gaaaataccg catcaggcgc tcatccgctt cctcgctcac tgactcgctg
     8101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     8161 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     8221 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     8281 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     8341 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     8401 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     8461 aggtatctca gttcggtgt
//
