LOCUS       sequence-391094-        3828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phytobrick (MoClo) Level 0 Part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3828)
  AUTHORS   Chamness JC, Kumar J, Cruz AJ, Rhuby E, Holum MJ, Cody JP, Tibebu R,
            Gamo ME, Starker CG, Zhang F, Voytas DF
  TITLE     An extensible vector toolkit and parts library for advanced 
            engineering of plant genomes.
  JOURNAL   Plant Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312.
  PUBMED    36896468
REFERENCE   2  (bases 1 to 3828)
  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 
            Genome. 2023 Mar 9:e20312. doi: 10.1002/tpg2.20312."
COMMENT     Sequence Label: pMC-6-GFP-P2A-NptII-9
FEATURES             Location/Qualifiers
     source          1..3828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(91..93,94..96,97..807)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(91..93,94..96,97..807)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(136..157)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(397..416)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     744..765
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             817..873
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             868..1668
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(928..947)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1538..1557
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1708..1726)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1794..1898
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2120..2911
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3082..3670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3571..3590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3824..3828,1..13)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
       61 gccgattcat taatcactct gtggtctcaa atggtgagca agggcgagga gctgttcacc
      121 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      181 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
      241 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
      301 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
      361 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
      421 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
      481 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
      541 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
      601 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
      661 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
      721 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
      781 actctcggca tggacgagct gtacaaggga agcggagcta ctaacttcag cctgctgaag
      841 caggctggtg acgtggagga gaaccctgga cctgggattg aacaagatgg attgcacgca
      901 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      961 ggctgctctg atgccgccgt gtttcggctg tcagcgcagg ggcgcccggt tctttttgtc
     1021 aagaccgacc tgtccggtgc cctgaatgaa ctccaggacg aggcagcgcg gctatcgtgg
     1081 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     1141 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     1201 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     1261 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     1321 gccggtcttg tcgatcagga tgatctggac gaagaacatc aggggctcgc gccagccgaa
     1381 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacacatggc
     1441 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1501 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1561 gaagaacttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1621 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc tttgagacca
     1681 cgaagtggct cttcagtgga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     1741 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1801 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1861 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gcgctcacgc aactggtcca
     1921 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     1981 actgtttttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     2041 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     2101 taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     2161 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     2221 cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc tggttacggt
     2281 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     2341 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     2401 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     2461 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     2521 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     2581 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     2641 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     2701 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     2761 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     2821 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc attacgtaaa
     2881 aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact catatatact
     2941 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3001 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3061 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3661 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3781 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcga
//
