LOCUS       sequence-391095-        3798 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 3798)
  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 3798)
  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-AmCyan-P2A-NptII-9
FEATURES             Location/Qualifiers
     source          1..3798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             91..777
                     /codon_start=1
                     /product="Anemonia majano cyan fluorescent protein"
                     /label=AmCyan1
                     /note="mammalian codon-optimized"
                     /translation="MALSNKFIGDDMKMTYHMDGCVNGHYFTVKGEGSGKPYEGTQTST
                     FKVTMANGGPLAFSFDILSTVFMYGNRCFTAYPTSMPDYFKQAFPDGMSYERTFTYEDG
                     GVATASWEISLKGNCFEHKSTFHGVNFPADGPVMAKMTTGWDPSFEKMTVCDGILKGDV
                     TAFLMLQGGGNYRCQFHTSYKTKKPVTMPPNHAVEHRIARTDLDKGGNSVQLTEHAVAH
                     ITSVVPF"
     CDS             787..843
                     /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             838..1638
                     /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(898..917)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1508..1527
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1678..1696)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1764..1868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2090..2881
                     /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      3052..3640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3541..3560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3794..3798,1..13)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
       61 gccgattcat taatcactct gtggtctcaa atggccctgt ccaacaagtt catcggcgac
      121 gacatgaaga tgacctacca catggacggc tgcgtgaacg gccactactt caccgtgaag
      181 ggcgagggca gcggcaagcc ctacgagggc acccagacct ccaccttcaa ggtgaccatg
      241 gccaacggcg gccccctggc cttctccttc gacatcctgt ccaccgtgtt catgtacggc
      301 aaccgctgct tcaccgccta ccccaccagc atgcccgact acttcaagca ggccttcccc
      361 gacggcatgt cctacgagag aaccttcacc tacgaggacg gcggcgtggc caccgccagc
      421 tgggagatca gcctgaaggg caactgcttc gagcacaagt ccaccttcca cggcgtgaac
      481 ttccccgccg acggccccgt gatggccaag atgaccaccg gctgggaccc ctccttcgag
      541 aagatgaccg tgtgcgacgg catcttgaag ggcgacgtga ccgccttcct gatgctgcag
      601 ggcggcggca actacagatg ccagttccac acctcctaca agaccaagaa gcccgtgacc
      661 atgcccccca accacgcggt ggagcaccgc atcgccagaa ccgacctgga caagggcggc
      721 aacagcgtgc agctcaccga gcacgccgtg gcccacatca cctccgtggt gcccttcgga
      781 agcggagcta ctaacttcag cctgctgaag caggctggtg acgtggagga gaaccctgga
      841 cctgggattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
      901 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gtttcggctg
      961 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     1021 ctccaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     1081 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     1141 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     1201 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     1261 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     1321 gaagaacatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     1381 gacggcgagg atctcgtcgt gacacatggc gatgcctgct tgccgaatat catggtggaa
     1441 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     1501 gacatagcgt tggctacccg tgatattgct gaagaacttg gcggcgaatg ggctgaccgc
     1561 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     1621 cttgacgagt tcttctgagc tttgagacca cgaagtggct cttcagtgga cgaaagggcc
     1681 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1741 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1801 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1861 ggaagagtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg
     1921 gcgcagtggc ggttttcatg gcttgttatg actgtttttt tttggggtac agtctatgcc
     1981 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     2041 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc
     2101 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     2161 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     2221 acacagcgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     2281 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     2341 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     2401 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     2461 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     2521 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     2581 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     2641 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     2701 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     2761 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     2821 tcagttggaa gaatttgtcc attacgtaaa aggcgagatc accaaggtag tcggcaaata
     2881 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3541 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3661 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3721 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3781 aacgaccgag cgcagcga
//
