LOCUS       sequence-343333-        4646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recoded sfGFP reporter with the codons TCG and TAG inserted at 
            positions 3 and 4, respectively..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4646)
  AUTHORS   Robertson WE, Funke LFH, de la Torre D, Fredens J, Elliott TS, 
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC, Chin JW
  TITLE     Sense codon reassignment enables viral resistance and encoded 
            polymer synthesis.
  JOURNAL   Science. 2021 Jun 4;372(6546):1057-1062. doi: 
            10.1126/science.abg3029.
  PUBMED    34083482
REFERENCE   2  (bases 1 to 4646)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2021-06-4- 4"; volume: "372"; issue: 
            "6546"; pages: "1057-1062"
COMMENT     Sequence Label: pBAD_sfGFP3TCG4TAG
FEATURES             Location/Qualifiers
     source          1..4646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..34)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      187..273
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(480..497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(614..1159)
                     /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(694..713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1271..1276,1277..1293,1294..1299)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        1347..1438
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2372..2394
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2550..3428)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3455..3739
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3662..3681
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3785..4495
                     /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="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4502..4519
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4545..4574
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4590..4607
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
       61 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
      121 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
      181 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
      241 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagctgtccc tcctgttcag
      301 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg
      361 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
      421 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
      481 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
      541 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
      601 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
      661 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
      721 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
      781 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
      841 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
      901 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
      961 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     1021 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     1081 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agtttacgcg
     1141 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc
     1201 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     1261 attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta tcacagttaa
     1321 attgctaacg cagtcaggca ccgtgttttg tttatttttc taaatacatt caaatatgta
     1381 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1441 gagtagtgcg gtggagtgca atgtcgtgca atacgaatgg cgaaaagccg agctcatcgg
     1501 tcagcttctc aaccttgggg ttacccccgg cggtgtgctg ctggtccaca gctccttccg
     1561 tagcgtccgg cccctcgaag atgggccact tggactgatc gaggccctgc gtgctgcgct
     1621 gggtccggga gggacgctcg tcatgcccag ctggagtggt ctggacgacg agccgttcga
     1681 tcctgccacg agccccgtta caccggacct tggagttgtc tctgacacat tctggcgcct
     1741 gccaaatgta aagcgcagcg cccatccatt tgcctttgcg gcagcggggc cacaggcaga
     1801 gcagatcatc tctgatccat tgcccctgcc acctcacagc cctgcaagcc cggtcgcccg
     1861 tgtccatgaa ctcgatgggc aggtacttct cctcggcgtg ggacacgatg ccaacacgac
     1921 gctgcatctt gccgagttga tggcaaaggt tccctatggg gtgccgagac actgcaccat
     1981 tcttcaggat ggcaagttgg tacgcgtcga ttatctcgag aatgaccact gctgtgagcg
     2041 ctttgccttg gcggacaggt ggctcaagga gaagagcctt cagaaggaag gtccagtcgg
     2101 tcatgccttt gctcggttga tccgctcccg cgacattgtg gcgacagccc tgggtcaact
     2161 gggccgagat ccgttgatct tcctgcatcc gccagaggcg ggatgcgaag aatgcgatgc
     2221 cgctcgccag agcattggct gaatggaagc cggcggcacc tcgctaacgg attcaccact
     2281 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg
     2341 cagaacatat ccatcgcgtg cggccgcgtc tccgggagct gcatgtgtca gaggttttca
     2401 ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat
     2461 gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc
     2521 aagccgtcaa ttgtctgatt cgttaccaat tagctcaact tgacggctac atcattcact
     2581 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
     2641 cggctgaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
     2701 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
     2761 acgctaatcc ctaactgctg gcggaaaaga tgactcagac ggctcgggct caagcaaaca
     2821 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
     2881 caagcctcgc gtacccgatt atccatcggt ggatggaggc tctcgttaat cgcttccatg
     2941 cgccgcagta acaattgctc aagcagattt atcgccagca gctcgctata gcgcccttcc
     3001 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
     3061 tccgggcgaa agaaccccgt attggcaaat atactcggcc agttaagcca ttcatgccag
     3121 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
     3181 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
     3241 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
     3301 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
     3361 aaacccgcca ccagatgggc attaaagctg tatcccggca gcaggggatc attttgcgct
     3421 tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc
     3481 agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg
     3541 cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa
     3601 agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc
     3661 tatgccatag catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa
     3721 ctctctactg tttctccata cccgtttttt gggctaacag gaggaattaa ccatggcttc
     3781 gtagagcaaa ggtgaagaac tgtttaccgg cgttgtgccg attctggtgg aactggatgg
     3841 tgatgtgaat ggccataaat ttagcgttcg tggcgaaggc gaaggtgatg cgaccaacgg
     3901 taaactgacc ctgaaattta tttgcaccac cggtaaactg ccggttccgt ggccgaccct
     3961 ggtgaccacc ctgacctatg gcgttcagtg ctttagccgc tatccggatc atatgaaacg
     4021 ccatgatttc tttaaaagcg cgatgccgga aggctatgtg caggaacgta ccattagctt
     4081 caaagatgat ggcacctata aaacccgtgc ggaagttaaa tttgaaggcg ataccctggt
     4141 gaaccgcatt gaactgaaag gtattgattt taaagaagat ggcaacattc tgggtcataa
     4201 actggaatat aatttcaaca gccataacgt gtatattacc gccgataaac agaaaaatgg
     4261 catcaaagcg aactttaaaa tccgtcacaa cgtggaagat ggtagcgtgc agctggcgga
     4321 tcattatcag cagaataccc cgattggtga tggcccggtg ctgctgccgg ataatcatta
     4381 tctgagcacc cagagcgttc tgagcaaaga tccgaatgaa aaacgtgatc atatggtgct
     4441 gctggaattt gttaccgccg cgggcattac ccacggtatg gatgaactgt ataaaggcag
     4501 ccaccatcat catcaccatt aaagctcgag cgaagcttgg gcccgaacaa aaactcatct
     4561 cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttaaacgg
     4621 tctccagctt ggctgttttg gcggat
//
