LOCUS       sequence-177670-        2602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  jStack yeast assembly into plant vectors, Level 1 Intermediary 
            Vector, ColE1origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2602)
  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 2602)
  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: pPMS008 (JBEI-11596)
FEATURES             Location/Qualifiers
     source          1..2602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             217..228
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             235..948
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(1102..1761)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1676..1695
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1943..2531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2432..2451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtag ggcgcgcctg caggcggccg cgaattggtc
       61 cagagaccga aagtgaaacg tgatttcatg cgtcattttg aacattttgt aaatcttatt
      121 taataatgtg tgcggcaatt cacatttaat ttatgaatgt tttcttaaca tcgcggcaac
      181 tcaagaaacg gcaggttcgg atcttagcta ctagagaaag aggagaaata ctagatgcgt
      241 aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc
      301 aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg
      361 acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg
      421 acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac
      481 ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat
      541 gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc
      601 attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa
      661 tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa
      721 gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac
      781 cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc
      841 acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag
      901 ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaatg aggtctctcg
      961 ctggatctcc tgcaggcggc cgcggcgcgc ctcaccaata ataacgcccg gcggcaaccg
     1021 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     1081 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     1141 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     1201 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     1261 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     1321 ggctgaaacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     1381 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     1441 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     1501 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     1561 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     1621 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     1681 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     1741 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     1801 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc
     1861 gatcaatcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2581 ttgctcacat gttctttcct gc
//
