LOCUS       sequence-314900-        2778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent protein CTAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2778)
  AUTHORS   Andreou , Andreas I., Nirkko, Jessica , Ochoa-Villarreal, Marisol , 
            Nakayama, Naomi
  TITLE     Mobius Assembly for Plant Systems highlights promoter-terminator 
            interaction in gene regulation
  JOURNAL   bioRxiv, March 31, 2021
REFERENCE   2  (bases 1 to 2778)
  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: "bioRxiv, 
            March 31, 2021"
COMMENT     Sequence Label: lvl0 sfGFP CTAG
FEATURES             Location/Qualifiers
     source          1..2778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      23..80
                     /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(104..121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(275..863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(355..374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1046..1140)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1161..1820)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1735..1754
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1982..2000
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2004..2047)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             join(2067..2069,2070..2778,1..5)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2109..2130)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2717..2738
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 agtaagcttt gagaccctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt
       61 tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc
      121 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      181 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      241 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca caggctccgc ccccctgacg
      301 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      481 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      601 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      661 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      721 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      781 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      841 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      901 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      961 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1021 cttggtctga cagctcgagg cttggattct caccaataaa aaacgcccgg cggcaaccga
     1081 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     1141 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     1201 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     1261 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     1321 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     1381 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     1441 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     1501 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     1561 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
     1621 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     1681 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     1741 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     1801 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     1861 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg
     1921 atcaactcga gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     1981 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     2041 tgatttctgg aattcggtct ctttcggtga gcaagggcga ggagctgttc accggggtgg
     2101 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgagaggcg
     2161 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca
     2221 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2281 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2341 acgtccagga gcgcaccatc tctttcaagg acgacggcac ttacaagacc cgcgccgagg
     2401 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2461 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata
     2521 tcactgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacgttg
     2581 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2641 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgttctgagc aaagacccca
     2701 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2761 gcatggacga gctgtaca
//
