LOCUS       sequence-316091-        6434 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 6434)
  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 6434)
  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_0616:Dendra
FEATURES             Location/Qualifiers
     source          1..6434
                     /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"
     CDS             2378..2401
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2658..3458
                     /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      3583..4171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4072..4091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgatagagtt tgtcctccct atcagtgata gataggctta aggagtaata cgactcacta
       61 tagggagaag gaggccatta tgaccggccc cgtcaacccc gacgaccggc gctcgttcag
      121 ctcgcgcacc ccggtcaacg agaaccccga cggcgtgcag taccgccgcg gtttcgtcac
      181 gcgacaccag gtgagcggct ggcgattcgt catgcgccgc atcgcttccg gcgtcgcgct
      241 gcacgacacc cgcatgctcg tcgacccgtt gcgcacgcag agccgcgccg tgctcaccgg
      301 tgcgctgatc ctcgtcacgg gtctggtcgg ctgcttcatc ttctcgctgt tccgtccggg
      361 cggcgtgccg ggcaacaacg cgatcctcgc cgaccggtcg acctcggcgc tgtacgtgcg
      421 ggtcggggaa caactgcacc cggtgctcaa cctgacctcg gcccgcctga tctccggatc
      481 accggacaac ccgacgatgg tcaagacctc cgagatcgac aagttcccgc gcggaaacct
      541 gctcggcatc cccggcgccc cggaacgcat ggtgcagaac gccgccaccg acgcggagtg
      601 gaccgtgtgt gacgccgtcg gcggcgccaa ccccggtgtg acagtgatcg ccggcccgct
      661 cggcgccgac ggggagcgtg ccgccccgct gccacccgac cacgcggtgc ttgtgcacag
      721 cgacgccgaa cccaaccccg gtgactggct gttgtgggac ggcaagcgca gcccgatcga
      781 cctggccgac cgcgccgtga ccgacgcgct gggcctcggc gggcaggccc tggcgccgag
      841 gcccatcgca gccgggctgt tcaacgcggt tcccgcggca cccgcgctga ccgcgccggt
      901 gatccccgac gccggtgccg caccgcaatt cgagctctcg ctgcccgtgc ccgtcggcgc
      961 ggtcgtggtg gcctacgacg ccgacaacac cgcgcggtac tacgccgtgt tgtccgatgg
     1021 gctgcagccc atatcgcccg tgctcgcggc gatcctgcgc aacaccgact cccacggttt
     1081 cgcgcagccg ccgcgactgg ggccggacga ggtggcccgt acgccgatgt cacgcggcct
     1141 ggacacctcg gcctatccgg acaaccccgt gaccctcgtc gaggcgtcgg cccatccggt
     1201 gacgtgcgcg cactggacga aaccgtctga cgccgcggag agttcgctgt cggtgctctc
     1261 aggcgcggtg ctgccgctgg ccgaaggcct gcacaccgtg gacctggtgg gtgcgggcgc
     1321 gggcggtgcg gccaaccgcg tcgcgctgac gcctggcacc ggatacttcg tccagacggt
     1381 cggagccgaa cccggctcgc ccaccgccgg atcgatgttc tgggtcagcg atacgggcgt
     1441 gcgctacggg atcgacacag ccgaggatga caaagtggtt gccgcactcg ggctttccac
     1501 ttcgccgctg ccggtgccgt ggtcggtgct gtcgcagttc gcggccggac ccgccctgtc
     1561 gcgcggcgat gcgctcgtcg cgcatgacgc cgtgtcgacc aacccgaact ctgcgcgtat
     1621 ggaggcctcc cgaaagcttg gtgcggccgc gggcaacacg cctggcatca atctgatcaa
     1681 ggaggacatg cgtgtcaagg tgcatatgga gggcaacgtg aacggtcacg ccttcgtgat
     1741 cgagggggag ggtaagggta agccttacga agggacccag accgccaacc tcaccgtcaa
     1801 ggagggcgcc cctctcccat tctcgtacga tatcctcact accgcggtgc attacgggaa
     1861 ccgcgtgttc accaagtacc ccgaggatat ccctgactac ttcaagcaga gcttcccgga
     1921 gggatactcc tgggagcgga cgatgacctt cgaagacaag ggtatctgta cgatccgcag
     1981 cgacatctcg ctggaagggg actgtttctt ccagaacgtc cgtttcaagg ggaccaactt
     2041 tccgccgaac ggcccggtca tgcagaagaa gaccctcaag tgggagccga gcacggagaa
     2101 gctgcacgtt cgggacggcc tcctggtcgg caacatcaac atggcgctgc tgctggaagg
     2161 cggcggccac tacctatgcg acttcaagac cacgtacaaa gcgaagaagg tcgtccagct
     2221 gcccgacgcg cacttcgtgg accaccgcat cgaaatcctg gggaacgatt cggactacaa
     2281 caaggtcaag ttgtacgagc acgctgtggc acgctatagc ccgctcccgt cgcaggtctg
     2341 gttcgaaggc ggcggcgggg agaacctgta tttccaggac tacaaggacg acgacgacaa
     2401 gtgatttaaa taaccccttg gggcctctaa acgggtcttg aggggttttt tgttaactag
     2461 cgtacgatcg aattcctgca gctcacggta actgatgccg tatttgcagt accagcgtac
     2521 ggcccacaga atgatgtcac gctgaaaatg ccggcctttg aatgggttca tgtgcagctc
     2581 catcagcaaa aggggatgat aagtttatca ccaccgacta tttgcaacag tgccgttgat
     2641 cgtgctatga tcgactgatg tcatcagcgg tggagtgcaa tgtcgtgcaa tacgaatggc
     2701 gaaaagccga gctcatcggt cagcttctca accttggggt tacccccggc ggtgtgctgc
     2761 tggtccacag ctccttccgt agcgtccggc ccctcgaaga tgggccactt ggactgatcg
     2821 aggccctgcg tgctacgctg ggtccgggag ggacgctcgt catgccctcg tggtcaggtc
     2881 tggacgacga gccgttcgat cctgccacgt cgcccgttac accggacctt ggagttgtct
     2941 ctgacacatt ctggcgcctg ccaaatgtaa agcgcagcgc ccatccattt gcctttgcgg
     3001 cagcggggcc acaggcagag cagatcatct ctgatccatt gcccctgcca ccttactcgc
     3061 ctgcaagccc ggtcgcccgt gtccatgaac tcgatgggca ggtacttctc ctcggcgtgg
     3121 gacacgatgc caacacgacg ctgcatcttg ccgagttgat ggcaaaggtt ccctatgggg
     3181 tgccgagaca ctgcaccatt cttcaggatg gcaagttggt acgcgtcgat tatctcgaga
     3241 atgaccactg ctgtgagcgc tttgccttgg cggacaggtg gctcaaggag aagagccttc
     3301 agaaggaagg tccagtcggt catgcctttg ctcggttgat ccgctcccgc gacattgtgg
     3361 cgacagccct gggtcaactg ggccgagatc cgttgatctt cctgcatccg ccagaggcgg
     3421 gatgcgaaga atgcgatgcc gctcgccagt cgattggctg agctcatgag cggagaacga
     3481 gatgacgttg gaggggcaag gtcgcgctga ttgctggggc aacacggggg atccactagt
     3541 tccatgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3721 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3901 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3961 cctacagcgt gagctatgag aaagcgccac gcttcccgag gggagaaagg cggacaggta
     4021 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4081 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4141 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4201 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4261 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4321 gcgcaacgcg tgcggccgcg gtacccgggg atcctctaga gtcgaccacc aagggcacca
     4381 tctctgcttg ggccaccccg ttggccgcag ccagctcgct gagagccgtg aacgacaggg
     4441 cgaacgccag cccgccgacg gcgagggttc cgaccgctgc aactcccggt gcaaccttgt
     4501 cccggtctat tctcttcact gcaccagctc caatctggtg tgaatgcccc tcgtctgttc
     4561 gcgcaggcgg ggggctctat tcgtttgtca gcatcgaaag tagccagatc agggatgcgt
     4621 tgcaaccgcg tatgcccagg tcagaagagt cgcacaagag ttgcagaccc ctggaaagaa
     4681 aaatggccag agggcgaaaa caccctctga ccagcggagc gggcgacggg aatcgaaccc
     4741 gcgtagctag tttggaagaa tgggtgtctg ccgaccacat atgggccggt caagataggt
     4801 ttttaccccc tctcggctgc atcctctaag tggaaagaaa ttgcaggtcg tagaagcgcg
     4861 ttgaagcctg agagttgcac aggagttgca acccggtagc cttgttcacg acgagaggag
     4921 acctagttgg cacgtcgcgg atggggatcg ctgaagactc agcgcagcgg gaggatccaa
     4981 gcctcatacg tcaacccgca ggacggtgtg aggtactacg cgctgcagac ctacgacaac
     5041 aagatggacg ccgaagcctg gctcgcgggc gagaagcggc tcatcgagat ggagacctgg
     5101 acccctccac aggaccgggc gaagaaggca gccgccagcg ccatcacgct ggaggagtac
     5161 acccggaagt ggctcgtgga gcgcgacctc gcagacggca ccagggatct gtacagcggg
     5221 cacgcggagc gccgcatcta cccggtgcta ggtgaagtgg cggtcacaga gatgacgcca
     5281 gctctggtgc gtgcgtggtg ggccgggatg ggtaggaagc acccgactgc ccgccggcat
     5341 gcctacaacg tcctccgggc ggtgatgaac acagcggtcg aggacaagct gatcgcagag
     5401 aacccgtgcc ggatcgagca gaaggcagcc gatgagcgcg acgtagaggc gctgacgcct
     5461 gaggagctgg acatcgtcgc cgctgagatc ttcgagcact accggatcgc ggcatacatc
     5521 ctggcgtgga cgagcctccg gttcggagag ctgatcgagc ttcgccgcaa ggacatcgtg
     5581 gacgacggca tgacgatgaa gctccgggtg cgccgtggcg cttcccgcgt ggggaacaag
     5641 atcgtcgttg gcaacgccaa gaccgtccgg tcgaagcgtc ctgtgacggt tccgcctcac
     5701 gtcgcggaga tgatccgagc gcacatgaag gaccgtacga agatgaacaa gggccccgag
     5761 gcattcctgg tgaccacgac gcagggcaac cggctgtcga agtccgcgtt caccaagtcg
     5821 ctgaagcgtg gctacgccaa gatcggtcgg ccggaactcc gcatccacga cctccgcgct
     5881 gtcggcgcta cgttcgccgc tcaggcaggt gcgacgacca aggagctgat ggcccgtctc
     5941 ggtcacacga ctcctaggat ggcgatgaag taccagatgg cgtctgaggc ccgcgacgag
     6001 gctatcgctg aggcgatgtc caagctggcc aagacctcct gaaacgcaaa aagcccccct
     6061 cccaaggaca ctgagtccta aagagggggg tttcttgtca gtacgcgaag aaccacgcct
     6121 ggccgcgagc gccagcaccg ccgctctgtg cggagacctg ggcaccagcc ccgccgccgc
     6181 caggagcatt gccgttcccg ccagaaatct agacggtgac cacaacgcgc ccgctttgat
     6241 cggggacgtc gaaatattgg atcgtcggca ccgtcacggc cgtgggaggc ggcacgatcc
     6301 gcgacgtgat gatcggccgc atccccacgg tgctgcgcag tgagctctac gccatcccgg
     6361 cgttgatctg tgcgttcgca cgcacaggcc cggtgtgaga agggtctctg acgagcggga
     6421 gaactcccta tcag
//
