LOCUS       sequence-275031-        2820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FP for C-term fusion  with CDS12 without linker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2820)
  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 2820)
  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_linker-eGFP
FEATURES             Location/Qualifiers
     source          1..2820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..87
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(91..134)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             join(193..195,196..198,199..912)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(193..195,196..198,199..909)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(238..259)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(499..518)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     846..867
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      930..987
                     /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(1011..1028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1182..1770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1262..1281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1953..2047)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2068..2727)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2642..2661
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtgcccgatc aactcgagtg ccacctgacg tctaagaaac cattattatc atgacattaa
       61 cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg
      121 ctaaggatga tttctggaat tcggtctcat tcgggagcag ctgcggctgc cgctgcggca
      181 gcggccggat ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      241 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      301 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      361 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      421 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      481 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      541 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      601 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
      661 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
      721 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      781 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
      841 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      901 ctgtacaagt aagcttcgag accctgcagt ccggcaaaaa agggcaaggt gtcaccaccc
      961 tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact
     1021 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1081 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1141 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag gctccgcccc
     1201 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1261 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1321 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1381 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1441 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1501 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1561 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1621 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1681 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1741 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1801 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1861 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1921 gagtaaactt ggtctgacag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg
     1981 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     2041 aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg
     2101 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     2161 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     2221 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     2281 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     2341 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     2401 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     2461 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg
     2521 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     2581 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     2641 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     2701 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     2761 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
//
