LOCUS       sequence-119083-        6968 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 6968)
  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 6968)
  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_alpha2R
FEATURES             Location/Qualifiers
     source          1..6968
                     /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    634..658
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1959..2588
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3022..4089
                     /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      4155..4349
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4585..4607)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4693..4833
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4848..4865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5019..5607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5099..5118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5695..6510)
                     /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(5695..6510)
                     /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     6418..6437
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6943..6967
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacgatatc gtctcagtca 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 ctctctccgc ttgagacgat atctgacagg atatattggc gggtaaacct
      661 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
      721 cgttcgtcca tttgtatgtg catgccaacc acagggttcc cctcgggatc aaagtacttt
      781 gatccaaccc ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcatc
      841 tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc
      901 ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg
      961 gagacattac gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag
     1021 caccgacgac caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa
     1081 gctgttttcc gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct
     1141 tgaccaccta cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag
     1201 cacccgcgac ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag
     1261 cctggcagag ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt
     1321 cgccggcatt gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga
     1381 ggccgccaag gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat
     1441 cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact
     1501 gcttggcgtg catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc
     1561 caccgaggcc aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct
     1621 ggcggccgcc gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag
     1681 gacgaaccgt ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg
     1741 ttcgagccgc ccgcgcacct ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct
     1801 gatgccaagc tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt
     1861 ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg
     1921 atgcgatgag taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta
     1981 accagaaagg cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac
     2041 tcgccggggc cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg
     2101 cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc
     2161 gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg
     2221 cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa
     2281 gcccttacga catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg
     2341 tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca
     2401 tcggcggtga ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta
     2461 tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag
     2521 aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac
     2581 tcatttgagt taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg
     2641 ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc
     2701 catgaagcgg gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc
     2761 ggtacgccaa ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga
     2821 gtaaatgagc aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg
     2881 aaaatcaaga acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg
     2941 gttggccagg cgtaagcggc tgggttgtct gccggccctg caatggcact ggaaccccca
     3001 agcccgagga atcggcgtga cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg
     3061 ctgggtgatg acctggtgga gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc
     3121 gaggcagaag cacgccccgg tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa
     3181 tcccggcaac cgccggcagc cggtgcgccg tcgattagga agccgcccaa gggcgacgag
     3241 caaccagatt ttttcgttcc gatgctctat gacgtgggca cccgcgatag tcgcagcatc
     3301 atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc
     3361 tacgagcttc cagacgggca cgtagaggtt tccgcagggc cggccggcat ggccagtgtg
     3421 tgggattacg acctggtact gatggcggtt tcccatctaa ccgaatccat gaaccgatac
     3481 cgggaaggga agggagacaa gcccggccgc gtgttccgtc cacacgttgc ggacgtactc
     3541 aagttctgcc ggcgagccga tggcggaaag cagaaagacg acctggtaga aacctgcatt
     3601 cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga aggccaagaa cggccgcctg
     3661 gtgacggtat ccgagggtga agccttgatt agccgctaca agatcgtaaa gagcgaaacc
     3721 gggcggccgg agtacatcga gatcgagcta gctgattgga tgtaccgcga gatcacagaa
     3781 ggcaagaacc cggacgtgct gacggttcac cccgattact ttttgatcga tcccggcatc
     3841 ggccgttttc tctaccgcct ggcacgccgc gccgcaggca aggcagaagc cagatggttg
     3901 ttcaagacga tctacgaacg cagtggcagc gccggagagt tcaagaagtt ctgtttcacc
     3961 gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg atttgaagga ggaggcgggg
     4021 caggctggcc cgatcctagt catgcgctac cgcaacctga tcgagggcga agcatccgcc
     4081 ggttcctaat gtacggagca gatgctaggg caaattgccc tagcagggga aaaaggtcga
     4141 aaaggactct ttcctgtgga tagcacgtac attgggaacc caaagccgta cattgggaac
     4201 cggaacccgt acattgggaa cccaaagccg tacattggga accggtcaca catgtaagtg
     4261 actgatataa aagagaaaaa aggcgatttt tccgcctaaa actctttaaa acttattaaa
     4321 actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc agcgcacagc cgaagagctg
     4381 caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct
     4441 atcgcggccg ctggccgctc aaaaatggct ggcctacggc caggcaatct accagggcgc
     4501 ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac atcaaggcac cctgcctcgc
     4561 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggtga cggtcacagc
     4621 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4681 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     4741 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     4801 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     4861 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4921 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4981 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5041 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5101 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5161 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5221 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5281 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5341 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5401 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5461 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5521 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5581 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5641 gctcagtgga acgaaaactc acgttaaggg attttggtca tgcattctag gtgattagaa
     5701 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     5761 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     5821 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     5881 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     5941 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     6001 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     6061 agcgagtcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     6121 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     6181 taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg catcatcagg
     6241 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     6301 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     6361 tggcgcatcg ggcttcccat acaatcggta gattgtcgca cctgattgcc cgacattatc
     6421 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggccttga
     6481 gcaagacgtt tcccgttgaa tatggctcat aacagaactt attatttcct tcctcttttc
     6541 tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt
     6601 tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt
     6661 tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa
     6721 cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc
     6781 cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct
     6841 tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga
     6901 ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg
     6961 tgtaaaca
//
