LOCUS       sequence-204789-        4577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used as a PCR template to amplify the glmS-HA fragment for 
            gene specific tagging and 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-glmS-4H
FEATURES             Location/Qualifiers
     source          1..4577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             701..781
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             1325..2350
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1325..2350
                     /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(2367..2383)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2367..2383)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2415..2434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2416..2434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2441..2458)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2441..2457)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2450..2472)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2598..3053)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2735..2756)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2947..2966
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3080..3184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3185..3253,3254..4045)
                     /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(3403..3422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4216..4577,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgaa attaaccctc
      661 actaaaggga acaaaagctg gtaccgggcc ccccctcgag tacccttacg atgtgcctga
      721 ttacgcgtac ccatacgacg tgccagacta cgcatacccg tacgatgtgc ccgattacgc
      781 atagtcgact gagtaattat agcgcccgaa ctaagcgccc ggaaaaaggc ttagttgacg
      841 aggatggagg ttatcgaatt ttcggcggat gcctcccggc tgagtgtgca gatcacagcc
      901 gtaaggattt cttcaaacca agggggtgac tccttgaaca aagagaaatc acatgatctg
      961 tcgacaaagt gtgacaacgt cgcaccatgt gtaggttttc atttatgttc tttctttctt
     1021 tcttttgtga atttgttttc tgtctcaaat gtttttaatt cgcttgggac ctatgttttt
     1081 cttgtttttt tgctcaccct ttgtgtagga ggcaccctgt cacgtctgtg gttgcgtgta
     1141 tgccttcctt ccccttattc gcttcttcct gtcgtgtcac acctctttct ccctctccct
     1201 ttccgccttt tctttcaatc ttgttttctc gaccagccct actagaggag aaagaatagt
     1261 aaccctttca tcaaagaaaa tagttcaaac gaattgctag cttaagcttg atctagaact
     1321 agtgatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     1381 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     1441 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     1501 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     1561 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     1621 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     1681 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     1741 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     1801 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     1861 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     1921 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     1981 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     2041 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     2101 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     2161 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     2221 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     2281 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     2341 aaaggaatag agtagatgcc gaccgggatc cactagttct agagcggccg ccaccgcggt
     2401 ggagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     2461 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2521 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2581 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2641 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2701 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2761 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2821 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2881 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2941 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3001 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3061 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3121 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3181 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3241 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3301 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3361 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3421 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3481 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3541 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3601 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3661 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3721 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3781 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3841 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3901 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3961 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4021 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4081 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4141 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4201 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4261 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4321 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4381 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4441 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4501 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4561 cttggagcga acgacct
//
