LOCUS       sequence-266748-        2741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sfGFP recoded in the RED20 genetic code off the T7 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2741)
  AUTHORS   Calles J, Justice I, Brinkley D, Garcia A, Endy D
  TITLE     Fail-safe genetic codes designed to intrinsically contain engineered
            organisms.
  JOURNAL   Nucleic Acids Res. 2019 Sep 12. pii: 5568210. doi: 
            10.1093/nar/gkz745.
  PUBMED    31511890
REFERENCE   2  (bases 1 to 2741)
  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: "Nucleic 
            Acids Res. 2019 Sep 12. pii: 5568210. doi: 10.1093/nar/gkz745."
COMMENT     Sequence Label: pSB1C3-pT7-sfGFP_RED20
FEATURES             Location/Qualifiers
     source          1..2741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(44..46,47..760)
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    786..806
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      807..864
                     /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(888..905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1059..1647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1139..1158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1830..1924)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1945..2604)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGIMLNELQQYCDEWQGGA"
     primer_bind     2519..2538
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 taatacgact cactataggg cttaagtata aggagatata catatgtcaa aaggagaaga
       61 actattcacc ggagtcgtcc cgatcctagt cgaactagac ggagacgtca acggacacaa
      121 attctcagtc cgtggagaag gagaaggaga cgcaaccaac ggaaaactaa ccctaaaatt
      181 catctgcacc accggaaaac taccggtccc gtggccgacc ctagtcacca ccctaaccta
      241 cggagtccag tgcttctcac gttacccgga ccacatgaaa cgtcacgact tcttcaaatc
      301 agcaatgccg gaaggatacg tccaggaacg taccatctca ttcaaagacg acggaaccta
      361 caaaacccgt gcagaagtca aattcgaagg agacacccta gtcaaccgta tcgaactaaa
      421 aggaatcgac ttcaaagaag acggaaacat cctaggacac aaactagaat acaacttcaa
      481 ctcacacaac gtctacatca ccgcagacaa acagaaaaac ggaatcaaag caaacttcaa
      541 aatccgtcac aacgtcgaag acggatcagt ccagctagca gaccactacc agcagaacac
      601 cccgatcgga gacggaccgg tcctactacc ggacaaccac tacctatcaa cccagtcagt
      661 cctatcaaaa gacccgaacg aaaaacgtga ccacatggtc ctactagaat tcgtcaccgc
      721 agcaggaatc acccacggaa tggacgaact atacaaatga cgctgatagt gctagtgtag
      781 atcgctacta gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc
      841 cctttttctt taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac
      901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccacaggct ccgcccccct
     1081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1381 gtaagacacg acttatcgca actggcagca gccactggta acaggattag cagagcgagg
     1441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1801 taaacttggt ctgacagctc gaggcttgga ttctcaccaa taaaaaacgc ccggcggcaa
     1861 ccgagcgttc tgaacaaatc cagatggagt tatgaggtca ttactggatc tatcaacagg
     1921 agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg
     1981 taattcatta agcattatgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
     2041 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     2101 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     2161 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     2221 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     2281 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     2341 aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc
     2401 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
     2461 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     2521 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     2581 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     2641 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     2701 cccgatcaac tcgagtgcca cctgacgtct aagaaaccga a
//
