LOCUS       sequence-119080-        6965 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 6965)
  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 6965)
  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_alpha1
FEATURES             Location/Qualifiers
     source          1..6965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     333..355
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     347..364
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     348..364
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    365..421
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(434..450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(434..450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(447..469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    458..474
                     /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)."
     promoter        complement(join(482..488,489..506,507..512))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    527..548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    631..655
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1956..2585
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3019..4086
                     /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      4152..4346
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4582..4604)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4690..4830
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4845..4862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5016..5604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5096..5115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5692..6507)
                     /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(5692..6507)
                     /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     6415..6434
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6940..6964
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacgaattc gtctcaggag agagacccac agcttgtctg taagcggatg ccgggagcag
       61 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      121 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
      181 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
      241 agggcgatcg gcgcaggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      301 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      361 cagtgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
      421 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      481 acaacatacg agccggaagc atatagtgta aagcctgggg tgcctaatga gtgagctaac
      541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      601 tggggtctct cgctgtcatg agacgaattc tgacaggata tattggcggg taaacctaag
      661 agaaaagagc gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt
      721 tcgtccattt gtatgtgcat gccaaccaca gggttcccct cgggatcaaa gtactttgat
      781 ccaacccctc cgctgctata gtgcagtcgg cttctgacgt tcagtgcagc cgtcatctga
      841 aaacgacatg tcgcacaagt cctaagttac gcgacaggct gccgccctgc ccttttcctg
      901 gcgttttctt gtcgcgtgtt ttagtcgcat aaagtagaat acttgcgact agaaccggag
      961 acattacgcc atgaacaaga gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac
     1021 cgacgaccag gacttgacca accaacgggc cgaactgcac gcggccggct gcaccaagct
     1081 gttttccgag aagatcaccg gcaccaggcg cgaccgcccg gagctggcca ggatgcttga
     1141 ccacctacgc cctggcgacg ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac
     1201 ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct
     1261 ggcagagccg tgggccgaca ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc
     1321 cggcattgcc gagttcgagc gttccctaat catcgaccgc acccggagcg ggcgcgaggc
     1381 cgccaaggcc cgaggcgtga agtttggccc ccgccctacc ctcaccccgg cacagatcgc
     1441 gcacgcccgc gagctgatcg accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct
     1501 tggcgtgcat cgctcgaccc tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac
     1561 cgaggccagg cggcgcggtg ccttccgtga ggacgcattg accgaggccg acgccctggc
     1621 ggccgccgag aatgaacgcc aagaggaaca agcatgaaac cgcaccagga cggccaggac
     1681 gaaccgtttt tcattaccga agagatcgag gcggagatga tcgcggccgg gtacgtgttc
     1741 gagccgcccg cgcacctctc aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat
     1801 gccaagctgg cggcctggcc ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta
     1861 aaaaggtgat gtgtatttga gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg
     1921 cgatgagtaa ataaacaaat acgcaagggg aacgcatgaa ggttatcgct gtacttaacc
     1981 agaaaggcgg gtcaggcaag acgaccatcg caacccatct agcccgcgcc ctgcaactcg
     2041 ccggggccga tgttctgtta gtcgattccg atccccaggg cagtgcccgc gattgggcgg
     2101 ccgtgcggga agatcaaccg ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg
     2161 acgtgaaggc catcggccgg cgcgacttcg tagtgatcga cggagcgccc caggcggcgg
     2221 acttggctgt gtccgcgatc aaggcagccg acttcgtgct gattccggtg cagccaagcc
     2281 cttacgacat atgggccacc gccgacctgg tggagctggt taagcagcgc attgaggtca
     2341 cggatggaag gctacaagcg gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg
     2401 gcggtgaggt tgccgaggcg ctggccgggt acgagctgcc cattcttgag tcccgtatca
     2461 cgcagcgcgt gagctaccca ggcactgccg ccgccggcac aaccgttctt gaatcagaac
     2521 ccgagggcga cgctgcccgc gaggtccagg cgctggccgc tgaaattaaa tcaaaactca
     2581 tttgagttaa tgaggtaaag agaaaatgag caaaagcaca aacacgctaa gtgccggccg
     2641 tccgagcgca cgcagcagca aggctgcaac gttggccagc ctggcagaca cgccagccat
     2701 gaagcgggtc aactttcagt tgccggcgga ggatcacacc aagctgaaga tgtacgcggt
     2761 acgccaaggc aagaccatta ccgagctgct atctgaatac atcgcgcagc taccagagta
     2821 aatgagcaaa tgaataaatg agtagatgaa ttttagcggc taaaggaggc ggcatggaaa
     2881 atcaagaaca accaggcacc gacgccgtgg aatgccccat gtgtggagga acgggcggtt
     2941 ggccaggcgt aagcggctgg gttgtctgcc ggccctgcaa tggcactgga acccccaagc
     3001 ccgaggaatc ggcgtgacgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg
     3061 ggtgatgacc tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag
     3121 gcagaagcac gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc
     3181 cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa
     3241 ccagattttt tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg
     3301 gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac
     3361 gagcttccag acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg
     3421 gattacgacc tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg
     3481 gaagggaagg gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag
     3541 ttctgccggc gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg
     3601 ttaaacacca cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg
     3661 acggtatccg agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg
     3721 cggccggagt acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc
     3781 aagaacccgg acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc
     3841 cgttttctct accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc
     3901 aagacgatct acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg
     3961 cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag
     4021 gctggcccga tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt
     4081 tcctaatgta cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa
     4141 ggactctttc ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg
     4201 aacccgtaca ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact
     4261 gatataaaag agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact
     4321 cttaaaaccc gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa
     4381 aaagcgccta cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc
     4441 gcggccgctg gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga
     4501 caagccgcgc cgtcgccact cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg
     4561 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggtgacgg tcacagcttg
     4621 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4681 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     4741 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     4801 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     4861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4921 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5641 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggtg attagaaaaa
     5701 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     5761 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     5821 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     5881 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     5941 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     6001 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     6061 gagtcgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     6121 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     6181 tacctggaat gctgttttcc ctgggatcgc agtggtgagt aaccatgcat catcaggagt
     6241 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     6301 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     6361 cgcatcgggc ttcccataca atcggtagat tgtcgcacct gattgcccga cattatcgcg
     6421 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gccttgagca
     6481 agacgtttcc cgttgaatat ggctcataac agaacttatt atttccttcc tcttttctac
     6541 agtatttaaa gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc
     6601 ttgcattcta aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg
     6661 cgtataacat agtatcgacg gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc
     6721 tctgtcatcg ttacaatcaa catgctaccc tccgcgagat catccgtgtt tcaaacccgg
     6781 cagcttagtt gccgttcttc cgaatagcat cggtaacatg agcaaagtct gccgccttac
     6841 aacggctctc ccgctgacgc cgtcccggac tgatgggctg cctgtatcga gtggtgattt
     6901 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt
     6961 aaaca
//
