LOCUS       sequence-204790-        4577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used as a PCR template to amplify the M9-HA point mutant of 
            glmS fragment for gene specific tagging that only allows tagging but
            not regulation. Acts as a control for glmS based gene regulation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4577)
  AUTHORS   Cruz-Bustos T, Ramakrishnan S, Cordeiro CD, Ahmed MA, Docampo R
  TITLE     A Riboswitch-based Inducible Gene Expression System for Trypanosoma 
            brucei.
  JOURNAL   J Eukaryot Microbiol. 2017 Dec 19. doi: 10.1111/jeu.12493.
  PUBMED    29265590
REFERENCE   2  (bases 1 to 4577)
  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: "J Eukaryot 
            Microbiol. 2017 Dec 19. doi: 10.1111/jeu.12493."
COMMENT     Sequence Label: pMOTag-M9-4H
FEATURES             Location/Qualifiers
     source          1..4577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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)."
     primer_bind     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        240..258
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             291..371
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             915..1940
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             915..1940
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(1957..1973)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1957..1973)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2005..2024)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2006..2024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2031..2048)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2031..2047)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2040..2062)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2188..2643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2325..2346)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2537..2556
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2670..2774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2775..2843,2844..3635)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2993..3012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3806..4394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4295..4314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4548..4565
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgaa
      241 attaaccctc actaaaggga acaaaagctg gtaccgggcc ccccctcgag tacccttacg
      301 atgtgcctga ttacgcgtac ccatacgacg tgccagacta cgcatacccg tacgatgtgc
      361 ccgattacgc atagtcgact gagtaattat cccgcccgaa ctaagcgccc ggaaaaaggc
      421 ttagttgacg aggatggagg ttatcgaatt ttcggcggat gcctcccggc tgagtgtgca
      481 gatcacagcc gtaaggattt cttcaaacca agggggtgac tccttgaaca aagagaaatc
      541 acatgatctg tcgacaaagt gtgacaacgt cgcaccatgt gtaggttttc atttatgttc
      601 tttctttctt tcttttgtga atttgttttc tgtctcaaat gtttttaatt cgcttgggac
      661 ctatgttttt cttgtttttt tgctcaccct ttgtgtagga ggcaccctgt cacgtctgtg
      721 gttgcgtgta tgccttcctt ccccttattc gcttcttcct gtcgtgtcac acctctttct
      781 ccctctccct ttccgccttt tctttcaatc ttgttttctc gaccagccct actagaggag
      841 aaagaatagt aaccctttca tcaaagaaaa tagttcaaac gaattgctag cttaagcttg
      901 atctagaact agtgatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga
      961 tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg
     1021 ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg
     1081 gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg
     1141 aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac
     1201 agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg
     1261 cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat
     1321 tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg
     1381 atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc
     1441 aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc
     1501 acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg
     1561 actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga
     1621 ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc
     1681 ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc
     1741 agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa
     1801 tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg
     1861 tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc
     1921 gtccgagggc aaaggaatag agtagatgcc gaccgggatc cactagttct agagcggccg
     1981 ccaccgcggt ggagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     2041 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     2101 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     2161 cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg
     2221 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2281 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2341 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2401 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2461 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2521 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2581 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2641 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2701 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2761 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2821 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2881 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2941 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3001 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3061 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3121 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3181 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3241 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3301 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3361 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3421 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3481 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3541 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3601 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3661 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3721 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3781 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3841 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3901 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3961 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4021 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4081 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4141 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4201 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4261 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4321 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4381 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4441 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4501 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4561 gcgaggaagc ggaagag
//
