LOCUS       sequence-317797-        2947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Erythromycin-dependent sfGFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2947)
  AUTHORS   Kolber NS, Fattal R, Bratulic S, Carver GD, Badran AH
  TITLE     Orthogonal translation enables heterologous ribosome engineering in 
            E. coli.
  JOURNAL   Nat Commun. 2021 Jan 26;12(1):599. doi: 10.1038/s41467-020-20759-z.
  PUBMED    33500394
REFERENCE   2  (bases 1 to 2947)
  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: "2021-01-26"; volume: "12(1):599. doi"; pages: " 
            10.1038/s41467-020-20759-z"
COMMENT     Sequence Label: pAB140j8
FEATURES             Location/Qualifiers
     source          1..2947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..33
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(106..123)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      276..362
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      454..481
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      525..1070
                     /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     971..990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1128..1943)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1128..1943)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1851..1870
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1935..1954
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    2101..2124
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 ggtattatgt ggagccatcc gcagttcgaa aaataagtcg accggctgct aacaaagccc
       61 gcggccgctg aagatcgatc tcgacgagtg agagaagatt ttcagcctga tacagattaa
      121 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
      181 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
      241 gtcaccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
      301 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
      361 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
      421 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
      481 ttgcgtttct acagagcgtc agacccctta ataagatgat cttcttgaga tcgttttggt
      541 ctgcgcgtaa tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag
      601 gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggaggagc gcagtcacca
      661 aaacttgtcc tttcagttta gccttaaccg gcgcatgact tcaagactaa ctcctctaaa
      721 tcaattacca gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt tggactcaag
      781 acgatagtta ccggataagg cgcagcggtc ggactgaacg gggggttcgt gcatacagtc
      841 cagcttggag cgaactgcct acccggaact gagtgtcagg cgtggaatga gacaaacgcg
      901 gccataacag cggaatgaca ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg
      961 agccgccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat
     1021 ttgagcgtca gatttcgtga tgcttgtcag gggggcggag cctatggaaa aacggctcaa
     1081 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca
     1141 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     1201 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     1261 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     1321 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     1381 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     1441 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     1501 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     1561 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     1621 tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     1681 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     1741 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     1801 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     1861 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac
     1921 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     1981 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     2041 acaacgtggc tttccatcaa aaaaatattg acaacataaa aaactttgtg ttatacttgt
     2101 ggaattgtga gcggataaca attctatatc tgttattttt tccaaccaca gatctatggg
     2161 catttttagt atttttgtaa tcagcacagt tcattatcaa ccaaacaaaa aattaagtgg
     2221 ttataatgaa tcgttaataa gcaaaattca ttataaccaa attagcaaag gtgaagaact
     2281 gtttaccggc gttgtgccga ttctggtgga actggatggc gatgtgaacg gtcacaaatt
     2341 cagcgtgcgt ggtgaaggtg aaggcgatgc cacgattggc aaactgacgc tgaaatttat
     2401 ctgcaccacc ggcaaactgc cggtgccgtg gccgacgctg gtgaccaccc tgacctatgg
     2461 cgttcagtgt tttagtcgct atccggatca catgaaacgt cacgatttct ttaaatctgc
     2521 aatgccggaa ggctatgtgc aggaacgtac gattagcttt aaagatgatg gcaaatataa
     2581 aacgcgcgcc gttgtgaaat ttgaaggcga taccctggtg aaccgcattg aactgaaagg
     2641 cacggatttt aaagaagatg gcaatatcct gggccataaa ctggaataca actttaatag
     2701 ccataatgtt tatattacgg cggataaaca gaaaaatggc atcaaagcga attttaccgt
     2761 tcgccataac gttgaagatg gcagtgtgca gctggcagat cattatcagc agaatacccc
     2821 gattggtgat ggtccggtgc tgctgccgga taatcattat ctgagcacgc agaccgttct
     2881 gtctaaagat ccgaacgaaa aaggcacgcg ggaccacatg gttctgcacg aatatgtgaa
     2941 tgcggca
//
