LOCUS       Exported                6967 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  GoldenBraid level 1 Alpha plant expression vector. .
ACCESSION   .
VERSION     .
KEYWORDS    pDGB3_alpha2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6967)
  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 6967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6967
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     338..360
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     352..369
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     353..369
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    370..426
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(439..455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(439..455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(452..474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    463..479
                     /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(487..517)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    532..553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    633..657
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1958..2587
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3021..4088
                     /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      4154..4348
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4584..4606)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4692..4832
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4847..4864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5018..5606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5098..5117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5694..6509)
                     /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     6417..6436
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    6942..6966
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 taacaagctt cgtctcagtc aggagagaga cccacagctt gtctgtaagc ggatgccggg
       61 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
      121 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
      181 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
      241 tgggaagggc gatcggcgca ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      301 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      361 acggccagtg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
      421 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
      481 tccacacaac atacgagccg gaagcatata gtgtaaagcc tggggtgcct aatgagtgag
      541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
      601 ccagctgggg tctctcgctt gagacgaagc tttgacagga tatattggcg ggtaaaccta
      661 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
      721 gttcgtccat ttgtatgtgc atgccaacca cagggttccc ctcgggatca aagtactttg
      781 atccaacccc tccgctgcta tagtgcagtc ggcttctgac gttcagtgca gccgtcatct
      841 gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct gcccttttcc
      901 tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga ctagaaccgg
      961 agacattacg ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc
     1021 accgacgacc aggacttgac caaccaacgg gccgaactgc acgcggccgg ctgcaccaag
     1081 ctgttttccg agaagatcac cggcaccagg cgcgaccgcc cggagctggc caggatgctt
     1141 gaccacctac gccctggcga cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc
     1201 acccgcgacc tactggacat tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc
     1261 ctggcagagc cgtgggccga caccaccacg ccggccggcc gcatggtgtt gaccgtgttc
     1321 gccggcattg ccgagttcga gcgttcccta atcatcgacc gcacccggag cgggcgcgag
     1381 gccgccaagg cccgaggcgt gaagtttggc ccccgcccta ccctcacccc ggcacagatc
     1441 gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg
     1501 cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc gcagcgagga agtgacgccc
     1561 accgaggcca ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc cgacgccctg
     1621 gcggccgccg agaatgaacg ccaagaggaa caagcatgaa accgcaccag gacggccagg
     1681 acgaaccgtt tttcattacc gaagagatcg aggcggagat gatcgcggcc gggtacgtgt
     1741 tcgagccgcc cgcgcacctc tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg
     1801 atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag cgccgccgtc
     1861 taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga
     1921 tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa
     1981 ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg ccctgcaact
     2041 cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc gcgattgggc
     2101 ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg
     2161 cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc
     2221 ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag
     2281 cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc gcattgaggt
     2341 cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat
     2401 cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg agtcccgtat
     2461 cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc ttgaatcaga
     2521 acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact
     2581 catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct aagtgccggc
     2641 cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga cacgccagcc
     2701 atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg
     2761 gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca gctaccagag
     2821 taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag gcggcatgga
     2881 aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg
     2941 ttggccaggc gtaagcggct gggttgtctg ccggccctgc aatggcactg gaacccccaa
     3001 gcccgaggaa tcggcgtgac ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc
     3061 tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg
     3121 aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat
     3181 cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc
     3241 aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca
     3301 tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct
     3361 acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt
     3421 gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc
     3481 gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca
     3541 agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc
     3601 ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg
     3661 tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg
     3721 ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag
     3781 gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg
     3841 gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt
     3901 tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg
     3961 tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc
     4021 aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg
     4081 gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa
     4141 aaggactctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc
     4201 ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga
     4261 ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa
     4321 ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc
     4381 aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta
     4441 tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg
     4501 gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg
     4561 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggtgac ggtcacagct
     4621 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4681 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     4741 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     4801 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     4861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     5461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcattctagg tgattagaaa
     5701 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     5761 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     5821 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     5881 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     5941 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     6001 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     6061 gcgagtcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     6121 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     6181 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     6241 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     6301 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     6361 ggcgcatcgg gcttcccata caatcggtag attgtcgcac ctgattgccc gacattatcg
     6421 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggccttgag
     6481 caagacgttt cccgttgaat atggctcata acagaactta ttatttcctt cctcttttct
     6541 acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca attcactgtt
     6601 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt
     6661 ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac
     6721 gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc
     6781 ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt
     6841 acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat
     6901 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt
     6961 gtaaaca
//
