LOCUS       sequence-177673-        2690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  jStack yeast assembly into plant vectors, Level 1 Intermediary 
            Vector, p15A origin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2690)
  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 2690)
  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: Ppms028(JBEI-11601)
FEATURES             Location/Qualifiers
     source          1..2690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             447..458
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     rep_origin      complement(912..1457)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(992..1011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1603..2262
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1669..1688)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2333..2367
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(2417..2690,1..440))
                     /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"
ORIGIN
        1 ctgttaaaat tgtattccag cttatggccc aggatattgc cgtcttcttt aaagtcaatg
       61 cctttcagct caatgcggtt taccagggta tcgccttcaa atttcacttc cgcacgcgtt
      121 ttgtacgtgc cgtcatcctt aaaggaaatc gtgcgttcct gcacatagcc ttccggcatg
      181 gcggacttga agaagtcatg ctgcttcata tggtccggat aacgagcaaa gcactgaaca
      241 ccataagtca gcgtcgttac cagagtcggc caaggtaccg gcagtttacc agtagtacag
      301 atgaacttca gcgtcagttt accattagtt gcgtcacctt caccctcgcc acgcacggaa
      361 aacttatgac cgttgacatc accatccagt tccaccagaa tagggacgac accagtgaac
      421 agctcttcgc ctttacgcat ctagtatttc tcctctttct ctagtagcta agatccgaac
      481 ctgccgtttc ttgagttgcc gcgatgttaa gaaaacattc ataaattaaa tgtgaattgc
      541 cgcacacatt attaaataag atttacaaaa tgttcaaaat gacgcatgaa atcacgtttc
      601 actttcggtc tctggaccaa ttcgcggccg cctgcaggcg cgccctagat ccggagtgta
      661 tactggctta ctatgttggc actgatgaga gtgtcagtga agtgcttcat gtggcaggag
      721 aaaaaaggct gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc
      781 gctcactggc tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg
      841 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
      901 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
      961 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     1021 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     1081 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     1141 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     1201 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     1261 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     1321 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     1381 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     1441 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc tcaattcttg
     1501 atcgggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     1561 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     1621 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     1681 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     1741 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     1801 atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat
     1861 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     1921 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     1981 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca
     2041 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     2101 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     2161 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     2221 gaattacaac agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct
     2281 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     2341 gctcggttgc cgccgggcgt tatttattgg tgaggcgcgc cgcggccgcc tgcaggagat
     2401 ccagcgagag acctcatttg tacagttcat ccataccatg cgtgatgccc gctgcggtta
     2461 cgaactccag cagaaccata tgatcgcgtt tctcgttcgg atctttagac agaacgcttt
     2521 gcgtgctcag atagtgattg tctggcagca gaacaggacc atcaccgatt ggagtgtttt
     2581 gctggtagtg atcagccagc tgcacgctgc catcctccac gttgtggcga attttaaaat
     2641 tcgctttaat gccatttttt tgtttatcgg cggtgatgta aacattgtgg
//
