LOCUS       sequence-307531-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses orthogonal gfp gene in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  AUTHORS   Aleksashin NA, Szal T, d'Aquino AE, Jewett MC, Vazquez-Laslop N, 
            Mankin AS
  TITLE     A fully orthogonal system for protein synthesis in bacterial cells.
  JOURNAL   Nat Commun. 2020 Apr 20;11(1):1858. doi: 10.1038/s41467-020-15756-1.
  PUBMED    32313034
REFERENCE   2  (bases 1 to 5668)
  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: "Nat 
            Commun."; date: "2020-04-20"; volume: "11(1):1858. doi"; pages: " 
            10.1038/s41467-020-15756-1"
COMMENT     Sequence Label: poGFP1
FEATURES             Location/Qualifiers
     source          1..5668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             63..74
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      878..949
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      965..992
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             1790..1813
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1886..1903)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2056..2142
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2234..2261
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        complement(join(2767..2772,2773..2789,2790..2795))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2907..3452
                     /direction=RIGHT
                     /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     3353..3372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3569..3586
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             4639..5430
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLAPRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcactactag
       61 agaaagagga gaaatactag atgaaaaaca taaatgccga cgacacatac agaataatta
      121 ataaaattaa agcttgtaga agcaataatg atattaatca atgcttatct gatatgacta
      181 aaatggtaca ttgtgaatat tatttactcg cgatcattta tcctcattct atggttaaat
      241 ctgatatttc aatcctagat aattacccta aaaaatggag gcaatattat gatgacgcta
      301 atttaataaa atatgatcct atagtagatt attctaactc caatcattca ccaattaatt
      361 ggaatatatt tgaaaacaat gctgtaaata aaaaatctcc aaatgtaatt aaagaagcga
      421 aaacatcagg tcttatcact gggtttagtt tccctattca tacggctaac aatggcttcg
      481 gaatgcttag ttttgcacat tcagaaaaag acaactatat agatagttta tttttacatg
      541 cgtgtatgaa cataccatta attgttcctt ctctagttga taattatcga aaaataaata
      601 tagcaaataa taaatcaaac aacgatttaa ccaaaagaga aaaagaatgt ttagcgtggg
      661 catgcgaagg aaaaagctct tgggatattt caaaaatatt aggttgcagt gagcgtactg
      721 tcactttcca tttaaccaat gcgcaaatga aactcaatac aacaaaccgc tgccaaagta
      781 tttctaaagc aattttaaca ggagcaattg attgcccata ctttaaaaat taataacact
      841 gatagtgcta gtgtagatca ctactagagc caggcatcaa ataaaacgaa aggctcagtc
      901 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
      961 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagacct gtaggatcgt
     1021 acaggtttac gcaagaaaat ggtttgttat agtcgaataa actatatctg ttattttttc
     1081 caaccacaga tctatgagca aaggtgaaga actgtttacc ggcgttgtgc cgattctggt
     1141 ggaactggat ggcgatgtga acggtcacaa attcagcgtg cgtggtgaag gtgaaggcga
     1201 tgccacgatt ggcaaactga cgctgaaatt tatctgcacc accggcaaac tgccggtgcc
     1261 gtggccgacg ctggtgacca ccctgaccta tggcgttcag tgttttagtc gctatccgga
     1321 tcacatgaaa cgtcacgatt tctttaaatc tgcaatgccg gaaggctatg tgcaggaacg
     1381 tacgattagc tttaaagatg atggcaaata taaaacgcgc gccgttgtga aatttgaagg
     1441 cgataccctg gtgaaccgca ttgaactgaa aggcacggat tttaaagaag atggcaatat
     1501 cctgggccat aaactggaat acaactttaa tagccataat gtttatatta cggcggataa
     1561 acagaaaaat ggcatcaaag cgaattttac cgttcgccat aacgttgaag atggcagtgt
     1621 gcagctggca gatcattatc agcagaatac cccgattggt gatggtccgg tgctgctgcc
     1681 ggataatcat tatctgagca cgcagaccgt tctgtctaaa gatccgaacg aaaaaggcac
     1741 gcgggaccac atggttctgc acgaatatgt gaatgcggca ggtattatgt ggagccatcc
     1801 gcagttcgaa aaataagtcg accggctgct aacaaagccc gcggccgctg aagatcgatc
     1861 tcgacgagtg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1921 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1981 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtcaccccat gcgagagtag
     2041 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2101 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     2161 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     2221 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acactcgata
     2281 ggggacgcca ctctaggaaa tccgccataa aacaaaaggc tcagtcggaa gactgggcct
     2341 tttgttttat gtcgacgggc ataaataggt ttaattttgc tacgggggcg ttatttaggt
     2401 tttttcttct ttcgaaaaaa tctttcttta tgaagttaaa agctatgtat tcaatagcat
     2461 attttgaata tggacataga atagtgctta tcactattgc atatagcatc ttatctgaca
     2521 caaggaaata atacccttcg ctgttttttg ttataaggta tatatatata agtgtgcagt
     2581 acaggccaaa taaaatattt tttatgtagt atcttaaatc ccgcaagagg cccggcagta
     2641 ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga
     2701 gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac
     2761 taccgcatta aagcttatcg atgataagct gtcaaacatg agaattacga cttatatcgt
     2821 atggggctga cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagaaatat
     2881 tttatctgat taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc
     2941 tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac
     3001 tctttgaacc gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt
     3061 tagccttaac cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc
     3121 tgccagtggt gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa
     3181 ggcgcagcgg tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc
     3241 ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga
     3301 caccggtaaa ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg
     3361 cctggtatct ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt
     3421 gatgcttgtc aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc
     3481 cctgttaagt atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc
     3541 cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat
     3601 cctgtatcac atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc
     3661 acttcactga caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct
     3721 gatgtccggc ggtgcttttg ccgttacgca ccaccccgtc agtagctgaa caggagggac
     3781 agagctttat gcttgtaaac cgttttgtga aaaaattttt aaaataaaaa aggggacctc
     3841 tagggtcccc aattaattag taatataatc tattaaaggt cattcaaaag gtcatccacc
     3901 ggatcaattc ccctgctcgc gcaggctggg tgccaagctc tcgggtaaca tcaaggcccg
     3961 atccttggag cccttgccct cccgcacgat gatcgtgccg tgatcgaaat ccagatcctt
     4021 gacccgcagt tgcaaaccct cactgatccg catggggccc ccgttccata cagaagctgg
     4081 gcgaacaaac gatgctcgcc ttccagaaaa ccgaggatgc gaaccacttc atccggggtc
     4141 agcaccaccg gcaagcgccg cgacggccga ggtcttccga tctcctgaag ccagggcaga
     4201 tccgtgcaca gcaccttgcc gtagaagaac agcaaggccg ccaatgcctg acgatgcgtg
     4261 gagaccgaaa ccttgcgctc gttcgccagc caggacagaa atgcctcgac ttcgctgctg
     4321 cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca
     4381 taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc
     4441 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     4501 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     4561 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
     4621 aaaacaaagt taaacatcat gagggaagcg gtgatcgccg aagtatcgac tcaactatca
     4681 gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac
     4741 ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg
     4801 accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg
     4861 gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac
     4921 gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc
     4981 aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg
     5041 ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt
     5101 gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac
     5161 tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg
     5221 tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag
     5281 cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa
     5341 gaagaagatc gcttggcccc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa
     5401 ggcgagatca ccaaggtagt cggcaaataa tgtctaacaa ttcgttcaag ccgacgccgc
     5461 ttcgcggcgc ggcttaactc aagcgttaga tgcactaagc acataattgc tcacagccaa
     5521 actatcaggt caagtctgct tttattattt ttaagcgtgc ataataagcc ctacacaaat
     5581 tgggagatat atcatgaaag gctggctttt tcttgttatc gcaatagttg gcgaagtaat
     5641 cgcaacatat tcgcggccgc ttctagag
//
