LOCUS       sequence-119082-        6969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid level 1 Alpha plant expression vector. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6969)
  AUTHORS   Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, 
            Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D
  TITLE     GoldenBraid 2.0: a comprehensive DNA assembly framework for plant 
            synthetic biology.
  JOURNAL   Plant Physiol. 2013 Jul;162(3):1618-31.
  PUBMED    23669743
REFERENCE   2  (bases 1 to 6969)
  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 
            Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31"
COMMENT     Sequence Label: pDGB3_alpha1R
FEATURES             Location/Qualifiers
     source          1..6969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    87..108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(123..128,129..146,147..153)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    161..177
                     /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)."
     primer_bind     166..188
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     185..201
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     185..201
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    214..270
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(271..288)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(271..287)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(280..302)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    635..659
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1960..2589
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3023..4090
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      4156..4350
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4586..4608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4694..4834
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4849..4866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5020..5608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5100..5119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5696..6511)
                     /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(5696..6511)
                     /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     6419..6438
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6944..6968
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacggatcc gtctcaggag agcgagagac cccagctggc acgacaggtt tcccgactgg
       61 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      121 gctttacact atatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      181 cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct
      241 agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
      301 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
      361 gcgtaatagc gaagaggcct gcgccgatcg cccttcccaa cagttgcgca gcctgaatgg
      421 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      481 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
      541 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      601 agctgtgggt ctctctccgt catgagacgg atcctgacag gatatattgg cgggtaaacc
      661 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
      721 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt
      781 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtcat
      841 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
      901 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
      961 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
     1021 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
     1081 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
     1141 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
     1201 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
     1261 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
     1321 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
     1381 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
     1441 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
     1501 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
     1561 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
     1621 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
     1681 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
     1741 gttcgagccg cccgcgcacc tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
     1801 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     1861 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     1921 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     1981 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
     2041 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
     2101 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
     2161 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
     2221 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
     2281 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
     2341 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
     2401 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
     2461 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
     2521 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
     2581 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     2641 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     2701 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     2761 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     2821 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     2881 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     2941 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
     3001 aagcccgagg aatcggcgtg acggtcgcaa accatccggc ccggtacaaa tcggcgcggc
     3061 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat
     3121 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga
     3181 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga
     3241 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat
     3301 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg
     3361 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt
     3421 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata
     3481 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact
     3541 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat
     3601 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct
     3661 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac
     3721 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga
     3781 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat
     3841 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt
     3901 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac
     3961 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg
     4021 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc
     4081 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg
     4141 aaaaggactc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa
     4201 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt
     4261 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa
     4321 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct
     4381 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc
     4441 tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg
     4501 cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg
     4561 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggtg acggtcacag
     4621 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4681 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     4741 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4801 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     4861 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4921 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5461 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgcattcta ggtgattaga
     5701 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     5761 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     5821 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     5881 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     5941 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     6001 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     6061 gagcgagtcg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     6121 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     6181 ctaatacctg gaatgctgtt ttccctggga tcgcagtggt gagtaaccat gcatcatcag
     6241 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     6301 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     6361 ctggcgcatc gggcttccca tacaatcggt agattgtcgc acctgattgc ccgacattat
     6421 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggccttg
     6481 agcaagacgt ttcccgttga atatggctca taacagaact tattatttcc ttcctctttt
     6541 ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc caattcactg
     6601 ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt gttttcaaag
     6661 ttggcgtata acatagtatc gacggagccg attttgaaac cgcggtgatc acaggcagca
     6721 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     6781 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
     6841 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
     6901 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     6961 gtgtaaaca
//
