LOCUS       sequence-260035-        8602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter of Agrobacterium rhizogenes mediated 
            transformed roots - modified.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8602)
  TITLE     Gresshoff/Ferguson plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8602)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pHairyRed (double 35s removed)
FEATURES             Location/Qualifiers
     source          1..8602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..56)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(39..55)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(48..70)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        432..777
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     720..740
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     intron          809..998
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             1054..1731
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(1234..1254)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1646..1669
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     terminator      1767..2019
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2041..2065
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3366..3995
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4424..5497
                     /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      5563..5757
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5993..6015)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6101..6241
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6156..6175
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6256..6273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6427..7015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6507..6526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7102..7896)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    8321..8345
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    8423..8597
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 gatcctctag agtcgacctg caggcatgca agcttggcac tggccgtcgt tttacaacgt
       61 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      121 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      181 ctgaatggcg aatgctagag cagcttgagc ttggatcaga ttgtcgtttc ccgccttcag
      241 tttagcttca tggagtcaaa gattcaaata gaggacctaa cagaactcgc cgtaaagact
      301 ggcgaacagt tcatacagag tctcttacga ctcaatgaca agaagaaaat cttcgtcaac
      361 atggtggagc acgacacact tgtctactcc aaaaatatca aagatacagt ctcagaagac
      421 caaagggcaa ttgagacttt tcaacaaagg gtaatatccg gaaacctcct cggattccat
      481 tgcccagcta tctgtcactt tattgtgaag atagtggaaa aggaaggtgg ctcctacaaa
      541 tgccatcatt gcgataaagg aaaggccatc gttgaagatg cctctgccga cagtggtccc
      601 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct
      661 tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac
      721 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gagaacacgg
      781 gggactcttg accatggtag atctgagggt aaatttctag tttttctcct tcattttctt
      841 ggttaggacc cttttctctt tttatttttt tgagctttga tctttcttta aactgatcta
      901 ttttttaatt gattggttat ggtgtaaata ttacatagct ttaactgata atctgattac
      961 tttatttcgt gtgtctatga tgatgatgat agttacagaa ccgacgaact agtaacggcc
     1021 gccagtgtgc tggaattcgc ccttcatgca actatggcct cctccgagaa cgtcatcacc
     1081 gagttcatgc gcttcaaggt gcgcatggag ggcaccgtga acggccacga gttcgagatc
     1141 gagggcgagg gcgagggccg cccctacgag ggccacaaca ccgtgaagct gaaggtgacc
     1201 aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca gtacggctcc
     1261 aaggtgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc cttccccgag
     1321 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggcgac cgtgacccag
     1381 gactcctccc tgcaggacgg ctgcttcatc tacaaggtga agttcatcgg cgtgaacttc
     1441 ccctccgacg gccccgtgat gcagaagaag accatgggct gggaggcctc caccgagcgc
     1501 ctgtaccccc gcgacggcgt gctgaagggc gagacccaca aggccctgaa gctgaaggac
     1561 ggcggccact acctggtgga gttcaagtcc atctacatgg ccaagaagcc cgtgcagctg
     1621 cccggctact actacgtgga cgccaagctg gacatcacct cccacaacga ggactacacc
     1681 atcgtggagc agtacgagcg caccgagggc cgccaccacc tgttcctgta gccacgtgtg
     1741 aattggtgac cagctcgaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa
     1801 gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta
     1861 agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta
     1921 gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg
     1981 ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattaaac tatcagtgtt
     2041 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
     2101 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
     2161 gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata gtgcagtcgg
     2221 cttctgacgt tcagtgcagc cgtcttctga aaacgacatg tcgcacaagt cctaagttac
     2281 gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt ttagtcgcat
     2341 aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga gcgccgccgc
     2401 tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca accaacgggc
     2461 cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg gcaccaggcg
     2521 cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg ttgtgacagt
     2581 gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat
     2641 ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc
     2701 ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc gttccctaat
     2761 catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga agtttggccc
     2821 ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg accaggaagg
     2881 ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc tgtaccgcgc
     2941 acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg ccttccgtga
     3001 ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc aagaggaaca
     3061 agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga agagatcgag
     3121 gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacgtctc aaccgtgcgg
     3181 ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc ggccagcttg
     3241 gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga gtaaaacagc
     3301 ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat acgcaagggg
     3361 aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag acgaccatcg
     3421 caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta gtcgattccg
     3481 atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg ctaaccgttg
     3541 tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg cgcgacttcg
     3601 tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc aaggcagccg
     3661 acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc gccgacctgg
     3721 tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg gcctttgtcg
     3781 tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg ctggccgggt
     3841 acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca ggcactgccg
     3901 ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc gaggtccagg
     3961 cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag agaaaatgag
     4021 caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca aggctgcaac
     4081 gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt tgccggcgga
     4141 ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta ccgagctgct
     4201 atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg agtagatgaa
     4261 ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc gacgccgtgg
     4321 aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg gttgcctgcc
     4381 ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgagcg gtcgcaaacc
     4441 atccggcccg gtacaaatcg gcgcggcgct gggtgatgac ctggtggaga agttgaaggc
     4501 cgcgcaggcc gcccagcggc aacgcatcga ggcagaagca cgccccggtg aatcgtggca
     4561 agcggccgct gatcgaatcc gcaaagaatc ccggcaaccg ccggcagccg gtgcgccgtc
     4621 gattaggaag ccgcccaagg gcgacgagca accagatttt ttcgttccga tgctctatga
     4681 cgtgggcacc cgcgatagtc gcagcatcat ggacgtggcc gttttccgtc tgtcgaagcg
     4741 tgaccgacga gctggcgagg tgatccgcta cgagcttcca gacgggcacg tagaggtttc
     4801 cgcagggccg gccggcatgg ccagtgtgtg ggattacgac ctggtactga tggcggtttc
     4861 ccatctaacc gaatccatga accgataccg ggaagggaag ggagacaagc ccggccgcgt
     4921 gttccgtcca cacgttgcgg acgtactcaa gttctgccgg cgagccgatg gcggaaagca
     4981 gaaagacgac ctggtagaaa cctgcattcg gttaaacacc acgcacgttg ccatgcagcg
     5041 tacgaagaag gccaagaacg gccgcctggt gacggtatcc gagggtgaag ccttgattag
     5101 ccgctacaag atcgtaaaga gcgaaaccgg gcggccggag tacatcgaga tcgagctagc
     5161 tgattggatg taccgcgaga tcacagaagg caagaacccg gacgtgctga cggttcaccc
     5221 cgattacttt ttgatcgatc ccggcatcgg ccgttttctc taccgcctgg cacgccgcgc
     5281 cgcaggcaag gcagaagcca gatggttgtt caagacgatc tacgaacgca gtggcagcgc
     5341 cggagagttc aagaagttct gtttcaccgt gcgcaagctg atcgggtcaa atgacctgcc
     5401 ggagtacgat ttgaaggagg aggcggggca ggctggcccg atcctagtca tgcgctaccg
     5461 caacctgatc gagggcgaag catccgccgg ttcctaatgt acggagcaga tgctagggca
     5521 aattgcccta gcaggggaaa aaggtcgaaa aggtctcttt cctgtggata gcacgtacat
     5581 tgggaaccca aagccgtaca ttgggaaccg gaacccgtac attgggaacc caaagccgta
     5641 cattgggaac cggtcacaca tgtaagtgac tgatataaaa gagaaaaaag gcgatttttc
     5701 cgcctaaaac tctttaaaac ttattaaaac tcttaaaacc cgcctggcct gtgcataact
     5761 gtctggccag cgcacagccg aagagctgca aaaagcgcct acccttcggt cgctgcgctc
     5821 cctacgcccc gccgcttcgc gtcggcctat cgcggccgct ggccgctcaa aaatggctgg
     5881 cctacggcca ggcaatctac cagggcgcgg acaagccgcg ccgtcgccac tcgaccgccg
     5941 gcgcccacat caaggcaccc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     6001 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     6061 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     6121 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     6181 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6841 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7081 tttggtcatg cattctaggt actaaaacaa ttcatccagt aaaatataat attttatttt
     7141 ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc
     7201 gatatcctcc ctgatcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc
     7261 gcttctccca agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc
     7321 tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc
     7381 atacagctcg cgcggatctt taaatggagt gtcttcttcc cagttttcgc aatccacatc
     7441 ggccagatcg ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt
     7501 atagggacaa tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc
     7561 gataatcttt tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc
     7621 ctcactcatg agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac
     7681 aggcagcttt ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt
     7741 ccctttatac cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt
     7801 atatacctta gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag
     7861 ttttttcaat tccggtgata ttctcatttt agccatttat tatttccttc ctcttttcta
     7921 cagtatttaa agatacccca agaagctaat tataacaaga cgaactccaa ttcactgttc
     7981 cttgcattct aaaaccttaa ataccagaaa acagcttttt caaagttgtt ttcaaagttg
     8041 gcgtataaca tagtatcgac ggagccgatt ttgaaaccgc ggtgatcaca ggcagcaacg
     8101 ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg
     8161 gcagcttagt tgccgttctt ccgaatagca tcggtaacat gagcaaagtc tgccgcctta
     8221 caacggctct cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt
     8281 ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg
     8341 taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta atgtactgaa
     8401 ttaacgccga attaattcgg gggatctgga ttttagtact ggattttggt tttaggaatt
     8461 agaaatttta ttgatagaag tattttacaa atacaaatac atactaaggg tttcttatat
     8521 gctcaacaca tgagcgaaac cctataggaa ccctaattcc cttatctggg aactactcac
     8581 acattattat ggagaaactc ga
//
