LOCUS       sequence-275029-        2814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FP with ER retention signal for C-term fusion with CDS12 with ER 
            targeting signal peptide. Contains Gly-His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2814)
  AUTHORS   Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M, 
            Markel K, Delmans M, West A, Patron NJ, Haseloff J
  TITLE     Systematic tools for reprogramming plant gene expression in a simple
            model, Marchantia polymorpha.
  JOURNAL   ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511.
  PUBMED    32163700
REFERENCE   2  (bases 1 to 2814)
  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: "ACS Synth 
            Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511."
COMMENT     Sequence Label: CTAG_eGFP-GHtag-HDEL
FEATURES             Location/Qualifiers
     source          1..2814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..71
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(75..118)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             join(138..140,141..851)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(138..140,141..851)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(180..201)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(441..460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     788..809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             861..878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      908..965
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(989..1006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1160..1748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1240..1259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1931..2025)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2046..2705)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2620..2639
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
       61 tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg
      121 gaattcggtc tcattcggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      181 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      241 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      301 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      361 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      421 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      481 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
      541 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
      601 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
      661 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
      721 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
      781 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
      841 agctgtacaa gggggggggg caccaccacc accaccacga tgagctttaa gcttcgagac
      901 cctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct ttaaaaccga
      961 aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1021 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1081 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1141 aggccgcgtt gctggcgttt ttccacaggc tccgcccccc tgacgagcat cacaaaaatc
     1201 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1261 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1321 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1381 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1441 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1501 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1561 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1621 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1681 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1741 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1861 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct
     1921 cgaggcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     1981 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata
     2041 tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     2101 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     2161 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     2221 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     2281 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     2341 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     2401 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     2461 agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga
     2521 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     2581 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     2641 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     2701 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     2761 gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa ctcg
//
