LOCUS       sequence-135585-        3192 bp    DNA     circular SYN 14-APR-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 3192)
  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 3192)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pDGB1_alpha1
FEATURES             Location/Qualifiers
     source          1..3192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     120..139
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     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"
     rep_origin      complement(746..1334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(826..845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1505..2320)
                     /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
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2228..2247
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2312..2331
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2611..3046
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    join(3173..3192,1..3)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
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 taaactaagt
      661 cgctgtatgt gtttgtttga gatctcatgt gagcaaaagg ccagcaaaag gccaggaacc
      721 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      781 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      841 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      901 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      961 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1021 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1081 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1141 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1201 tctgcgctct gctgaagcca gttaccttcg gaagaagagt tggtagctct tgatccggca
     1261 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1321 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1381 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1441 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgtgtaa cattggtcta
     1501 gtgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1561 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1621 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1681 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1741 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     1801 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1861 attgcgcctg agcgagtcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     1921 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     1981 gatattcttc taatacctgg aatgctgttt tccctgggat cgcagtggtg agtaaccatg
     2041 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2101 agtttagtct gaccatctca tctgtaacaa cattggcaac gctacctttg ccatgtttca
     2161 gaaacaactc tggcgcatcg ggcttcccat acaatcggta gattgtcgca cctgattgcc
     2221 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2281 gcggccttga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2341 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     2401 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     2461 aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca
     2521 aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc ctcactttct
     2581 ggctggatga tggggcgatt caggcgatcc ccatccaaca gcccgccgtc gagcgggctt
     2641 ttttatcccc ggaagcctgt ggatagaggg tagttatcca cgtgaaaccg ctaatgcccc
     2701 gcaaagcctt gattcacggg gctttccggc ccgctccaaa aactatccac gtgaaatcgc
     2761 taatcagggt acgtgaaatc gctaatcgga gtacgtgaaa tcgctaataa ggtcacgtga
     2821 aatcgctaat caaaaaggca cgtgagaacg ctaatagccc tttcagatca acagcttgca
     2881 aacacccctc gctccggcaa gtagttacag caagtagtat gttcaattag cttttcaatt
     2941 atgaatatat atatcaatta ttggtcgccc ttggcttgtg gacaatgcgc tacgcgcacc
     3001 ggctccgccc gtggacaacc gcaagcggtt gcccaccgtc gagcgccttt gcccacaacc
     3061 cggcggccgg ccgcaacaga tcgttttata aatttttttt tttgaaaaag aaaaagcccg
     3121 aaaggcggca acctctcggg cttctggatt tccgatcccc ggaattagat cttggcagga
     3181 tatattgtgg tg
//
