LOCUS       sequence-368900-        7214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry under the virB promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  AUTHORS   Aliu E, Lee K, Wang K
  TITLE     CRISPR RNA-guided integrase enables high-efficiency targeted genome 
            engineering in Agrobacterium tumefaciens.
  JOURNAL   Plant Biotechnol J. 2022 Jun 11. doi: 10.1111/pbi.13872.
  PUBMED    35690588
REFERENCE   2  (bases 1 to 7214)
  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: "Plant 
            Biotechnol J. 2022 Jun 11. doi: 10.1111/pbi.13872."
COMMENT     Sequence Label: pEA106
FEATURES             Location/Qualifiers
     source          1..7214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             10..21
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             28..732
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMEELY"
     primer_bind     441..460
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      757..828
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(829..856)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2106..2735
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3164..4237
                     /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      4303..4497
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4733..4755)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4841..4981
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4896..4915
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4996..5013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5167..5755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5247..5266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6001..6795)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MAREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKP
                     HSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPA
                     KRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDL
                     FEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQ
                     PVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gaattcatta aagaggagaa aacgcgtatg gtgagcaagg gcgaggagga taacatggcc
       61 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      121 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      181 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
      241 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
      301 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
      361 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
      421 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
      481 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
      541 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
      601 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
      661 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
      721 gaggagctgt acagatctta aaagctttaa ggatctcaaa taaaacgaaa ggctcagtcg
      781 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctccg cagaaaggcc
      841 cacccgaagg tgagccgcat gccaaccaca gggttcccct cgggatcaaa gtactttaaa
      901 gtactttaaa gtactttaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg
      961 cttctgacgt tcagtgcagc cgtcttctga aaacgacatg tcgcacaagt cctaagttac
     1021 gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat
     1081 aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc
     1141 tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc
     1201 cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg
     1261 cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt
     1321 gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat
     1381 ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc
     1441 ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat
     1501 catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc
     1561 ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg
     1621 ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc
     1681 acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga
     1741 ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca
     1801 agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag
     1861 gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacgtctc aaccgtgcgg
     1921 ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg
     1981 gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc
     2041 ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg
     2101 aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg
     2161 caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg
     2221 atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg
     2281 tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg
     2341 tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg
     2401 acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg
     2461 tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg
     2521 tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt
     2581 acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg
     2641 ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg
     2701 cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag
     2761 caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac
     2821 gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga
     2881 ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct
     2941 atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa
     3001 ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg
     3061 aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgtctgcc
     3121 ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc
     3181 atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc
     3241 cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca
     3301 agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc
     3361 gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga
     3421 cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg
     3481 tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc
     3541 cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc
     3601 ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt
     3661 gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca
     3721 gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg
     3781 tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag
     3841 ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc
     3901 tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc
     3961 cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc
     4021 cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc
     4081 cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc
     4141 ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg
     4201 caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca
     4261 aattgcccta gcaggggaaa aaggtcgaaa aggtctcttt cctgtggata gcacgtacat
     4321 tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta
     4381 cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc
     4441 cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact
     4501 gtctggccag cgcacagccg aagagctgca aaaagcgcct acccttcggt cgctgcgctc
     4561 cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg
     4621 cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg
     4681 gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     4741 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     4801 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     4861 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     4921 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4981 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5041 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5101 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5161 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5221 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5281 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5341 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5401 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5461 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5521 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5581 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     5641 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5701 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5761 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5821 tttggtcatg catgatatat ctcccaattt gtgtagggct tattatgcac gcttaaaaat
     5881 aataaaagca gacttgacct gatagtttgg ctgtgagcaa ttatgtgctt agtgcatcta
     5941 acgcttgagt taagccgcgc cgcgaagcgg cgtcggcttg aacgaatttc tagctagaca
     6001 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
     6061 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
     6121 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     6181 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     6241 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     6301 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     6361 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgattgtggc
     6421 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     6481 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     6541 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     6601 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     6661 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
     6721 gagatggcgc tcgatgacgc caactacctc tgatagttga gttgatactt cggcgataac
     6781 cgcttcacga gccatggatc cctccctaca actagtgata tcgcccggag ttcgctccca
     6841 gaaatatagc catccctgcc ccctcacgtc aatcccacga gcctcttatc cattctcatt
     6901 gaacgacggc gaattgtatt gattggtgag ctaatcaata tcaaatatga aagggaaatc
     6961 gttaaattat tcaattatta tctaattgaa taatttaaga ataatggtac ctccggttgc
     7021 ttagtcatgg atgcgcggtt ctcggtccat gttgcgttcc aagacgccgg gcgaggtttc
     7081 tcgcttcaat tgaaatcata aagaagcaat tgaaaatttt cgagtaaccg accctcccga
     7141 taatcttcaa cataaaacaa cgcacttctt ccaacgggag aggcggtgtt agttgcgagc
     7201 taaggagata aggt
//
