LOCUS       sequence-316089-        6230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 expression of C-term fluorescent fusion in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6230)
  AUTHORS   Judd JA, Canestrari J, Clark R, Joseph A, Lapierre P, 
            Lasek-Nesselquist E, Mir M, Palumbo M, Smith C, Stone M, Upadhyay A,
            Wirth SE, Dedrick RM, Meier CG, Russell DA, Dills A, Dove E, Kester 
            J, Wolf ID, Zhu J, Rubin ER, Fortune S, Hatfull GF, Gray TA, Wade 
            JT, Derbyshire KM
  TITLE     A Mycobacterial Systems Resource for the Research Community.
  JOURNAL   mBio. 2021 Mar 2;12(2). pii: mBio.02401-20. doi: 
            10.1128/mBio.02401-20.
  PUBMED    33653882
REFERENCE   2  (bases 1 to 6230)
  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: "mBio."; 
            date: "2021-03-2"; volume: "12(2). pii: mBio.02401-20. doi"; pages: 
            " 10.1128/mBio.02401-20"
COMMENT     Sequence Label: pMSR:MSMEG_0372:Dendra
FEATURES             Location/Qualifiers
     source          1..6230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     46..65
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        46..64
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      919..928
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2174..2197
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2454..3254
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRATLGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPYSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      3379..3967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3868..3887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgatagagtt tgtcctccct atcagtgata gataggctta aggagtaata cgactcacta
       61 tagggagaag gaggccatta tggcttcgga cctgttttcc caagtggtca attccgggcc
      121 gggatcgttc ctggccaagc agctgggcgt gccgcagccc gagacgttgc gccgctaccg
      181 ccccggtgac ccgccgctgg cgggctcgct gctgatcggc ggatccggcc gcgtggccga
      241 acccctgcgc accgcattgg ccgacgacta caacctggtg tccaacaaca tcggtggccg
      301 gtgggccgat tcgttcggcg gcgtggtgtt cgacgccacg ggcatcaccg aggccgaggg
      361 cctcaaggag ctctacacgt tcttcacgcc gctgctgcgc aacctcgcgc cgtgtgcgcg
      421 ggtggtcgtc gtcggcacca cgcccgcgga ggccggcagc gtccacgcac aggtggtgca
      481 gcgggcgctg gaaggtttca cgcgctcgct gggcaaggag ctgcgccgtg gcgcgaccgt
      541 ctccctggtc tatctctcgg ccgacgccaa gccgggcgcc acgggtctcg agtcgaccat
      601 gcggttcatc ctgtcggcca agtcggccta cgtcgacggt caggtgttcc gcgtcggcgc
      661 cgccgattcg actccgcccg ccgactggga caagccgctc gacggcaagg tcgccgtggt
      721 caccggcgcg gcccgcggta tcggcgcgac gatcgccgag gtgttcgcgc gtgacggcgc
      781 gaccgtggtg gcgatcgacg tcgacggtgc ggccgaggat ctcaagcgcg tcgccgacaa
      841 ggtgggcggc accgcactca cgctcgacgt gaccgccgac gacgccgtcg acaagatcac
      901 cgcgcacgtc accgaacacc atggcggcaa ggtcgacatc ctcgtcaaca acgcgggtat
      961 cacgcgcgac aagctgctcg ccaacatgga cgagaagcgc tgggacgcgg tgatcgccgt
     1021 caacctgctc gcgccgcagc gcctcaccga aggcctggtg ggcaacggaa ccatcggcga
     1081 gggcggccgg gtgatcggcc tgtcgtcgat ggcgggcatc gcgggcaacc gcggccagac
     1141 caattacgcc accaccaagg ccgggatgat cgggctggcc gaggcgctgg cgcccgtgct
     1201 ggccgacaag ggcatcacga tcaacgccgt cgcacccggc ttcatcgaga ccaagatgac
     1261 cgaggccatc ccgctcgcca cgcgtgaggt gggcaggcgg ctcaactcgc tgttccaggg
     1321 cggccagccc gtcgacgtgg ccgagttgat cgcgtacttc gccagccccg cctcgaacgc
     1381 ggtgacgggc aacaccatcc gcgtgtgcgg ccaggcgatg ctgggggcga agcttggtgc
     1441 ggccgcgggc aacacgcctg gcatcaatct gatcaaggag gacatgcgtg tcaaggtgca
     1501 tatggagggc aacgtgaacg gtcacgcctt cgtgatcgag ggggagggta agggtaagcc
     1561 ttacgaaggg acccagaccg ccaacctcac cgtcaaggag ggcgcccctc tcccattctc
     1621 gtacgatatc ctcactaccg cggtgcatta cgggaaccgc gtgttcacca agtaccccga
     1681 ggatatccct gactacttca agcagagctt cccggaggga tactcctggg agcggacgat
     1741 gaccttcgaa gacaagggta tctgtacgat ccgcagcgac atctcgctgg aaggggactg
     1801 tttcttccag aacgtccgtt tcaaggggac caactttccg ccgaacggcc cggtcatgca
     1861 gaagaagacc ctcaagtggg agccgagcac ggagaagctg cacgttcggg acggcctcct
     1921 ggtcggcaac atcaacatgg cgctgctgct ggaaggcggc ggccactacc tatgcgactt
     1981 caagaccacg tacaaagcga agaaggtcgt ccagctgccc gacgcgcact tcgtggacca
     2041 ccgcatcgaa atcctgggga acgattcgga ctacaacaag gtcaagttgt acgagcacgc
     2101 tgtggcacgc tatagcccgc tcccgtcgca ggtctggttc gaaggcggcg gcggggagaa
     2161 cctgtatttc caggactaca aggacgacga cgacaagtga tttaaataac cccttggggc
     2221 ctctaaacgg gtcttgaggg gttttttgtt aactagcgta cgatcgaatt cctgcagctc
     2281 acggtaactg atgccgtatt tgcagtacca gcgtacggcc cacagaatga tgtcacgctg
     2341 aaaatgccgg cctttgaatg ggttcatgtg cagctccatc agcaaaaggg gatgataagt
     2401 ttatcaccac cgactatttg caacagtgcc gttgatcgtg ctatgatcga ctgatgtcat
     2461 cagcggtgga gtgcaatgtc gtgcaatacg aatggcgaaa agccgagctc atcggtcagc
     2521 ttctcaacct tggggttacc cccggcggtg tgctgctggt ccacagctcc ttccgtagcg
     2581 tccggcccct cgaagatggg ccacttggac tgatcgaggc cctgcgtgct acgctgggtc
     2641 cgggagggac gctcgtcatg ccctcgtggt caggtctgga cgacgagccg ttcgatcctg
     2701 ccacgtcgcc cgttacaccg gaccttggag ttgtctctga cacattctgg cgcctgccaa
     2761 atgtaaagcg cagcgcccat ccatttgcct ttgcggcagc ggggccacag gcagagcaga
     2821 tcatctctga tccattgccc ctgccacctt actcgcctgc aagcccggtc gcccgtgtcc
     2881 atgaactcga tgggcaggta cttctcctcg gcgtgggaca cgatgccaac acgacgctgc
     2941 atcttgccga gttgatggca aaggttccct atggggtgcc gagacactgc accattcttc
     3001 aggatggcaa gttggtacgc gtcgattatc tcgagaatga ccactgctgt gagcgctttg
     3061 ccttggcgga caggtggctc aaggagaaga gccttcagaa ggaaggtcca gtcggtcatg
     3121 cctttgctcg gttgatccgc tcccgcgaca ttgtggcgac agccctgggt caactgggcc
     3181 gagatccgtt gatcttcctg catccgccag aggcgggatg cgaagaatgc gatgccgctc
     3241 gccagtcgat tggctgagct catgagcgga gaacgagatg acgttggagg ggcaaggtcg
     3301 cgctgattgc tggggcaaca cgggggatcc actagttcca tgagcgtcag accccgtaga
     3361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3481 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3781 cgccacgctt cccgagggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4021 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4081 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc aacgcgtgcg gccgcggtac
     4141 ccggggatcc tctagagtcg accaccaagg gcaccatctc tgcttgggcc accccgttgg
     4201 ccgcagccag ctcgctgaga gccgtgaacg acagggcgaa cgccagcccg ccgacggcga
     4261 gggttccgac cgctgcaact cccggtgcaa ccttgtcccg gtctattctc ttcactgcac
     4321 cagctccaat ctggtgtgaa tgcccctcgt ctgttcgcgc aggcgggggg ctctattcgt
     4381 ttgtcagcat cgaaagtagc cagatcaggg atgcgttgca accgcgtatg cccaggtcag
     4441 aagagtcgca caagagttgc agacccctgg aaagaaaaat ggccagaggg cgaaaacacc
     4501 ctctgaccag cggagcgggc gacgggaatc gaacccgcgt agctagtttg gaagaatggg
     4561 tgtctgccga ccacatatgg gccggtcaag ataggttttt accccctctc ggctgcatcc
     4621 tctaagtgga aagaaattgc aggtcgtaga agcgcgttga agcctgagag ttgcacagga
     4681 gttgcaaccc ggtagccttg ttcacgacga gaggagacct agttggcacg tcgcggatgg
     4741 ggatcgctga agactcagcg cagcgggagg atccaagcct catacgtcaa cccgcaggac
     4801 ggtgtgaggt actacgcgct gcagacctac gacaacaaga tggacgccga agcctggctc
     4861 gcgggcgaga agcggctcat cgagatggag acctggaccc ctccacagga ccgggcgaag
     4921 aaggcagccg ccagcgccat cacgctggag gagtacaccc ggaagtggct cgtggagcgc
     4981 gacctcgcag acggcaccag ggatctgtac agcgggcacg cggagcgccg catctacccg
     5041 gtgctaggtg aagtggcggt cacagagatg acgccagctc tggtgcgtgc gtggtgggcc
     5101 gggatgggta ggaagcaccc gactgcccgc cggcatgcct acaacgtcct ccgggcggtg
     5161 atgaacacag cggtcgagga caagctgatc gcagagaacc cgtgccggat cgagcagaag
     5221 gcagccgatg agcgcgacgt agaggcgctg acgcctgagg agctggacat cgtcgccgct
     5281 gagatcttcg agcactaccg gatcgcggca tacatcctgg cgtggacgag cctccggttc
     5341 ggagagctga tcgagcttcg ccgcaaggac atcgtggacg acggcatgac gatgaagctc
     5401 cgggtgcgcc gtggcgcttc ccgcgtgggg aacaagatcg tcgttggcaa cgccaagacc
     5461 gtccggtcga agcgtcctgt gacggttccg cctcacgtcg cggagatgat ccgagcgcac
     5521 atgaaggacc gtacgaagat gaacaagggc cccgaggcat tcctggtgac cacgacgcag
     5581 ggcaaccggc tgtcgaagtc cgcgttcacc aagtcgctga agcgtggcta cgccaagatc
     5641 ggtcggccgg aactccgcat ccacgacctc cgcgctgtcg gcgctacgtt cgccgctcag
     5701 gcaggtgcga cgaccaagga gctgatggcc cgtctcggtc acacgactcc taggatggcg
     5761 atgaagtacc agatggcgtc tgaggcccgc gacgaggcta tcgctgaggc gatgtccaag
     5821 ctggccaaga cctcctgaaa cgcaaaaagc ccccctccca aggacactga gtcctaaaga
     5881 ggggggtttc ttgtcagtac gcgaagaacc acgcctggcc gcgagcgcca gcaccgccgc
     5941 tctgtgcgga gacctgggca ccagccccgc cgccgccagg agcattgccg ttcccgccag
     6001 aaatctagac ggtgaccaca acgcgcccgc tttgatcggg gacgtcgaaa tattggatcg
     6061 tcggcaccgt cacggccgtg ggaggcggca cgatccgcga cgtgatgatc ggccgcatcc
     6121 ccacggtgct gcgcagtgag ctctacgcca tcccggcgtt gatctgtgcg ttcgcacgca
     6181 caggcccggt gtgagaaggg tctctgacga gcgggagaac tccctatcag
//
