LOCUS       sequence-355677-        5731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Trypanosoma brucei PCR only tagging (pPOT) vector for endogenous 
            gene tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5731)
  AUTHORS   Paterou A, Saez Conde J, Tyc J, Sunter JD, Vaughan S, Gull K, Dean S
  TITLE     A comprehensive toolkit for protein localization and functional 
            analysis in trypanosomatids.
  JOURNAL   Open Biol. 2025 Apr;15(4):240361. doi: 10.1098/rsob.240361. Epub 
            2025 Apr 2.
  PUBMED    40169015
REFERENCE   2  (bases 1 to 5731)
  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: "Open 
            Biol."; date: "2025-04"; volume: "15(4):240361. doi"; pages: " 
            10.1098/rsob.240361. Epub 2025 Apr 2"
COMMENT     Sequence Label: pPOTv6-blast-3Myc:mNG:3Myc
FEATURES             Location/Qualifiers
     source          1..5731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             954..983
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             990..1019
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1026..1055
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1773..1802
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1809..1838
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1845..1874
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2856..2874)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2857..2874)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2892..2908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2892..2908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2905..2927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2916..2932
                     /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)."
     promoter        complement(join(2940..2946,2947..2964,2965..2970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2985..3006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3123..3140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3294..3882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3374..3393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4053..4844,4845..4913))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVIIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4676..4695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4914..5018)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5096..5551)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5233..5254)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5445..5464
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5677..5699
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5691..5708
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5692..5708
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5715..5731,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5715..5731,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgtata atgcagacct gctgcaagct aggggcacag caaggtcttc
       61 tgaaattcat gttttttttt tttttactct gcattgcagt ctccgctctt atttagtttt
      121 gctttacgta aggtctcgtt gctgccataa aataagctct agtggaattc atgcctttgt
      181 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
      241 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
      301 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
      361 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
      421 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga cctgcatcct gggatcaaag
      481 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
      541 ctggttatgt gtgggagggc taaccatggt gcccatttag ttggcctttc ccttgtctcg
      601 tgtcttttcc gtggaaaggt tcccggagta atctgatggc acagcaggga ggtgcgcctg
      661 caggttggtt aggaaggggg gatgatgtaa aagaagaaaa tggggggatt cgagccccac
      721 gcgttgctca agctgtgtag cgcaggcgtt tccttacata tttctctaac aggcacggaa
      781 gcctaacaaa tacacttggc ttattttttt gccccctcat gtcttgtaca aatatttgcg
      841 atagcttagc tatcagccac attaatcaaa caagtatacc aacaagcccg aaaacagaaa
      901 ctcaactgca acgggtacct atgggttctg gtagtggttc cggttccggt tctgagcaga
      961 agctgataag tgaagaagac ttgggctccg agcaaaagct gatttcggaa gaggatctgg
     1021 gctctgagca gaagctgatt tctgaagagg atcttaagct tgtctcgaaa ggtgaggaag
     1081 acaatatggc cagcttgcca gcaacgcacg agcttcatat atttgggtcg atcaacggtg
     1141 tggatttcga catggtgggt caaggcacag gtaatcctaa tgacgggtat gaagaactca
     1201 atttgaagag cactaaggga gatctccagt tttccccgtg gattcttgtg ccgcatattg
     1261 gttatggttt tcatcagtac ttaccatacc ccgatggaat gtcgcccttc caagcagcta
     1321 tggtggacgg ctcggggtat caagtacatc gcaccatgca atttgaggac ggagcatccc
     1381 tcaccgtaaa ctaccgctat acgtatgagg ggagccacat aaaaggtgag gctcaagtta
     1441 aaggcacagg cttcccagct gacggcccgg tcatgaccaa ttcattaacc gctgctgatt
     1501 ggtgccgttc taagaagacc taccctaacg acaagacgat cattagtact tttaagtgga
     1561 gttataccac aggaaacgga aagcggtatc gaagcactgc tcgaacaacc tacacatttg
     1621 cgaagccaat ggccgcgaac tacttgaaga accagccaat gtatgtgttt cgaaaaacgg
     1681 aactgaagca ttcgaaaacc gagcttaatt ttaaagagtg gcagaaggcg tttacagacg
     1741 tgatggggat ggacgaattg tataagggat ccgagcagaa gctgataagc gaggaagacc
     1801 tgggtagtga acagaagctg atctcagaag aggaccttgg ctcagaacaa aaactcatct
     1861 cagaggaaga tcttggatca ggatctggat caggatcggg tagttaagag ctccgctgcg
     1921 cttaaatgtc tttcattata atcaatgtat aacctttatg tagtatttca atctatgccg
     1981 ctgtgtacgt gcactgcggt gcctatcctt cggcattaga gagtcactgt ttgtgtagat
     2041 cgtagctgca tgtctgacat ctggttcatt agtgccgttt tgagtctgca ctgttgtgca
     2101 acttgatatt catcactgca caccaaccgt gtgtttcctc ctttgggtgc accctttaat
     2161 ccttgtcttc tccttttttg tctctttccc cctcgaaaag gtgtcaaact actgccgcat
     2221 aaactacggt tatcccaaat ttaagagaaa gcaataaagc atcagctagc atgcctttgt
     2281 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     2341 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     2401 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     2461 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     2521 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     2581 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     2641 ctggttatgt gtgggagggc taactcgagt gcccatttag ttggcttttc ccttgtctcg
     2701 tgtcttttcc gtggaaaggt tcccggagta atctgatggc acagcaggga ggtgcgcctg
     2761 caggttggtt aggaaggggg gatgatgtaa aagaagaaaa tggggggatt cgagccccgc
     2821 acaggtctct caaattggat gcatagcttg agtattctat agtgtcacct aaatagcttg
     2881 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2941 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3001 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3061 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3121 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3481 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3721 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3901 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3961 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4021 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4081 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4141 attacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4201 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4261 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4321 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4381 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4441 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4501 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4561 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4621 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4681 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4741 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4801 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4861 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4921 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4981 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5041 cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     5101 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5161 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5221 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5281 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5341 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5401 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5461 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5521 gccgcactta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     5581 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5641 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5701 cggccagtga attgtaatac gactcactat a
//
