LOCUS       sequence-332740-        4064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chromosomally-Integrated Fluorescent Mycobacterium tuberculosis 
            Reporter Constructs for mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4064)
  AUTHORS   Kolbe K, Bell AC, Prosser GA, Assmann M, Yang HJ, Forbes HE, 
            Gallucci S, Mayer-Barber KD, Boshoff HI, Barry Iii CE
  TITLE     Development and Optimization of Chromosomally-Integrated Fluorescent
            Mycobacterium tuberculosis Reporter Constructs.
  JOURNAL   Front Microbiol. 2020 Dec 9;11:591866. doi: 
            10.3389/fmicb.2020.591866. eCollection 2020.
  PUBMED    33362741
REFERENCE   2  (bases 1 to 4064)
  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: "Front 
            Microbiol."; date: "2020-12-9"; volume: "11:591866. doi"; pages: " 
            10.3389/fmicb.2020.591866. eCollection 2020"
COMMENT     Sequence Label: Plasmid L5 attB::Pleft* mCherry
FEATURES             Location/Qualifiers
     source          1..4064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             245..253
                     /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)"
     CDS             269..286
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1065..1111
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1920..1939)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1931..2746
                     /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
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2004..2023)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3077..3665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3566..3585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3895..3914
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3895..3913
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4030..4047
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatacgcaaa ccgcctctcc cagatctgat atcgctagag gaaacagcta tgaccatgat
       61 tacgccaagc ttgcatgcct gcagctaggg caccaatttg cgattagggc ttgacagcca
      121 cccggccagt agtgcattct tgtgtcaccg cagcagcaag gcggtaggcg gatccgagag
      181 gatcgtgccg gtgccggtga agagagattc gccgcccgaa atgagcacga tccgcatgct
      241 taattaagga ggtatctcca tgagcggtca ccatcaccat caccacggcg gtagcgccat
      301 gtctgtctcg aagggcgagg aggacaacat ggcgatcatc aaggagttca tgcgcttcaa
      361 ggtccacatg gagggctcgg tcaacggcca cgagttcgag atcgagggcg agggcgaggg
      421 ccgcccgtac gagggcaccc agaccgccaa gctgaaggtc accaagggcg gcccgctgcc
      481 gttcgcctgg gacatcctgt cgccgcagtt catgtacggc agcaaggcct acgtcaagca
      541 cccggccgac atcccggact acctgaagct gtcgttcccg gagggcttca agtgggagcg
      601 cgtcatgaac ttcgaggacg gcggcgtcgt caccgtcacc caggactcgt cgctgcagga
      661 cggcgagttc atctacaagg tcaagctgcg gggcaccaac ttcccgtcgg acggcccggt
      721 catgcagaag aagaccatgg gctgggaggc ctcgtcggag cgcatgtacc cggaggacgg
      781 cgccctgaag ggcgagatca agcagcggct gaagctgaag gacggcggcc actacgacgc
      841 cgaggtcaag accacctaca aggccaagaa gccggtccag ctgccgggcg cctacaacgt
      901 gaacatcaag ctggacatca ccagccacaa cgaggactac accatcgtcg agcagtacga
      961 gcgcgccgag ggccgccaca gcaccggcgg catggacgag ctgtacaagt gaaagcttat
     1021 cgatgtcgac gtagttaact agcgtacgat cgactgccag gcatcaaata aaacgaaagg
     1081 ctcagtcgaa agactgggcc tttcgtttta tgccatcatg gccgcggtga tcagctagag
     1141 ccgtgaacga cagggcgaac gccagcccgc cgacggcgag ggttccgacc gctgcaactc
     1201 ccggtgcaac cttgtcccgg tctattctct tcactgcacc agctccaatc tggtgtgaat
     1261 gcccctcgtc tgttcgcgca ggcggggggc tctattcgtt tgtcagcatc gaaagtagcc
     1321 agatcaggga tgcgttgcaa ccgcgtatgc ccaggtcaga agagtcgcac aagagttgca
     1381 gacccctgga aagaaaaatg gccagagggc gaaaacaccc tctgaccagc ggagcgggcg
     1441 acgggaatcg aacccgcgta gctagtttgg aagaatgggt gtctgccgac cacatatatg
     1501 ggccggtcaa gataggtttt taccccctct cggctgcatc ctctaagtgg aaagaaattg
     1561 caggtcgtag aagcgcgttg aagcctgaga gttgcacagg agttgcaacc cggtagcctt
     1621 gttcacgacg agaggagacc tagggatcct cccaaggaca ctgagtccta aagagggggg
     1681 tttcttgtca gtacgcgaag aaccacgcct ggccgcgagc gccagcaccg ccgctctgtg
     1741 cggagacctg ggcaccagcc ccgccgccgc caggagcatt gccgttcccg ccagctagcc
     1801 acctgacgtc gggggggggg aaagccacgt tgtgtctcaa aatctctgat gttacattgc
     1861 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1921 aaggggtgtt atgagccata ttcaacggga aacgtcttgc tcgaggccgc gattaaattc
     1981 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2041 tgcgacaatc tatcgcttgt atgggaagcc ccatgcgcca gagttgtttc tgaaacatgg
     2101 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2161 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2221 caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2281 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2341 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2401 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2461 agtctggaaa gaaatgcata atcttttgcc attctcaccg gattcagtcg tcactcatgg
     2521 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2581 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2641 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2701 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2761 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct ttgttgaata
     2821 aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt
     2881 tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa agctctcacc
     2941 aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg gtatgagtca
     3001 gcaacacctt cttcacgagg cagacctcac tagttccatg agcgtcagac cccgtagaaa
     3061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3481 ccacgcttcc cgaggggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3781 gagctgatac cgctcgccgc agccgaacga ccgagcgcaa cgcgtgcggc cgcggtacca
     3841 gatctttaaa tctagacgat gataagcggt caaacatgag aattcgcggc cgcataatac
     3901 gactcactat agggatctta attaaggcgc ctcatgttct ttcctgcgtt atcccctgat
     3961 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4021 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gccc
//
