LOCUS       sequence-323502-        4382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of green fluorescent protein encoding zero 
            stop codons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4382)
  AUTHORS   Ho JML, Miller CA, Parks SE, Mattia JR, Bennett MR
  TITLE     A suppressor tRNA-mediated feedforward loop eliminates leaky gene 
            expression in bacteria.
  JOURNAL   Nucleic Acids Res. 2021 Mar 18;49(5):e25. doi: 10.1093/nar/gkaa1179.
  PUBMED    33290521
REFERENCE   2  (bases 1 to 4382)
  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."; date: "2021-03-18"; volume: "49(5):e25. doi"; pages: " 
            10.1093/nar/gkaa1179"
COMMENT     Sequence Label: pJH805_pJH_LldR_Pcon-sfGFP.0TAG
FEATURES             Location/Qualifiers
     source          1..4382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(59..127,128..919)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(277..296)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1090..1678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1579..1598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1768..1785)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1938..2024
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2116..2143
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2445..3158
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             3451..3459
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      4297..4344
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
       61 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      121 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      181 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      241 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      301 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      361 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      421 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      481 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      541 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      601 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      661 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      721 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      781 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      841 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      901 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      961 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1021 caaaatccct taacgtgggt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1081 aggatcatct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1201 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcatggctg
     1741 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     1801 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     1861 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     1921 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1981 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     2041 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     2101 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     2161 tgctagcaag taaggccgac caacaaccta agggcaattc tctgatgagg attgcccttt
     2221 tctttaccag acatctcccc ccacaagctt cgcttatctg acctctggtt cacaatttcc
     2281 caattaaaac tcacatcaat gttgccaata cataacattt agttaaccat tcattgtcat
     2341 tatccctaca caacacttac acaacgtgtg acaaggagat gagcaacaga ctcattacac
     2401 gatgtgcgtg gactcccatt taactttaag aaggagatat acatatgcgt aaaggtgaag
     2461 aactgttcac cggtgttgtt ccgatcctgg ttgaactgga cggtgacgtt aacggtcaca
     2521 aattctctgt tcgtggtgaa ggtgaaggtg acgctaccaa cggtaaactg accctgaaat
     2581 tcatctgcac caccggtaaa ctgccggttc cgtggccgac cctggttacc accctgacct
     2641 acggtgttca gtgcttcgct cgttacccgg accacatgaa acagcacgac ttcttcaaat
     2701 ctgctatgcc ggaaggttac gttcaggaac gtaccatctc tttcaaagac gacggtacct
     2761 acaaaacccg tgctgaagtt aaattcgaag gtgacaccct ggttaaccgt atcgaactga
     2821 aaggtatcga cttcaaagaa gacggtaaca tcctgggtca caaactggaa tacaacttca
     2881 actctcacaa cgtttacatc accgctgaca aacagaaaaa cggtatcaaa gctaacttca
     2941 aaatccgtca caacgttgaa gacggttctg ttcagctggc tgaccactac cagcagaaca
     3001 ccccgatcgg tgacggtccg gttctgctgc cggacaacca ctacctgtct acccagtctg
     3061 ttctgtctaa agacccgaac gaaaaacgtg accacatggt tctgctggaa ttcgttaccg
     3121 ctgctggtat cacccacggt atggacgaac tgtacaaaga tgcatgccag ttctaacata
     3181 accctaatga gtgagctaac ttacattaat tgcgttgcgc cttaattaac ggcactcctc
     3241 agcaaatata atgaccctct tgataaccca agagggcatt ttttaatgcc catggcgttt
     3301 accacagcta acaccacgtc gtccctatct gctgccctag gtctatgagt ggttgctgga
     3361 taactttacg ggcatgcata aggctcgtag gctatattca gggagaccac aacggtttcc
     3421 ctctacaaat aattttgttt aactttgaaa taaggaggta atacaaatga ttgttttacc
     3481 cagacgcctg tcagacgagg ttgccgatcg tgtgcgggcg ctgattgatg aaaaaaacct
     3541 ggaagcgggc atgaagttgc ccgctgagcg ccaactggcg atgcaactcg gcgtatcacg
     3601 taattcactg cgcgaggcgc tggcaaaact ggtgagtgaa ggcgtgctgc tcagtcgacg
     3661 cggcggcggg acgtttattc gctggcgtca tgacacatgg tcggagcaaa acatcgtcca
     3721 gccgctaaaa acactgatgg ccgatgatcc ggattacagt ttcgatattc tggaagcccg
     3781 ctacgccatt gaagccagca ccgcatggca tgcggcaatg cgcgccacac ctggcgacaa
     3841 agaaaagatt cagctttgct ttgaagcaac gctaagtgaa gacccggata tcgcctcaca
     3901 agcggacgtt cgttttcatc tggcgattgc cgaagcctca cataacatcg tgctgctgca
     3961 aaccatgcgc ggtttcttcg atgtcctgca atcctcagtg aagcatagcc gtcagcggat
     4021 gtatctggtg ccaccggttt tttcacaact gaccgaacaa catcaggctg tcattgacgc
     4081 catttttgcc ggtgatgctg acggggcgcg taaagcaatg atggcgcacc ttagttttgt
     4141 tcacaccacc atgaaacgat tcgatgaaga tcaggctcgc cacgcacgga ttacccgcct
     4201 gcccggtgag cataatgagc attcgaggga gaaaaacgca taataagcga ctaaaaaatt
     4261 gaatgtagga aaccaacatg ccagttcgag caataactag cataacccct tggggcctct
     4321 aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca
     4381 ct
//
