LOCUS       sequence-177614-        2335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  jStack yeast assembly into plant vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2335)
  AUTHORS   Shih PM, Vuu K, Mansoori N, Ayad L, Louie KB, Bowen BP, Northen TR, 
            Loque D
  TITLE     A robust gene-stacking method utilizing yeast assembly for plant 
            synthetic biology.
  JOURNAL   Nat Commun. 2016 Oct 26;7:13215. doi: 10.1038/ncomms13215.
  PUBMED    27782150
REFERENCE   2  (bases 1 to 2335)
  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: "Nat 
            Commun."; date: "2016-10-26"; volume: "7:13215. doi"; pages: " 
            10.1038/ncomms13215"
COMMENT     Sequence Label: pms5092(JBEI-14430)
FEATURES             Location/Qualifiers
     source          1..2335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(77..868,869..937))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     700..719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(938..1042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1110..1128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1132..1175)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     terminator      1196..1444
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     primer_bind     complement(1482..1499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1653..2241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1733..1752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
       61 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      181 gcttaccatc tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag
      241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      361 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
      421 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      481 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
      541 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
      601 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
      661 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
      721 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
      781 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
      841 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
      901 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
      961 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1021 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     1081 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat
     1141 aaaaaaaatc cttatctttc gctaaggatg atttctggaa ttcggtctct gcttatatga
     1201 agatgaagat gaaatatttg gtgtgtcaaa taaaaagctt gtgtgcttaa gtttgtgttt
     1261 ttttcttggc ttgttgtgtt atgaatttgt ggctttttct aatattaaat gaatgtaaga
     1321 tctcattata atgaataaac aaatgtttct ataatccatt gtgaatgttt tgttggatct
     1381 cttctgcagc atataactac tgtatgtgct atggtatgga ctatggaata tgattaaaga
     1441 taagcgcttg agaccggatc ctaactcgag ctgcaggctt cctcgctcac tgactcgctg
     1501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1621 aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag
     1681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatct
//
