LOCUS       sequence-355471-        5722 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 5722)
  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 5722)
  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:mScarletI:3Myc
FEATURES             Location/Qualifiers
     source          1..5722
                     /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             1764..1793
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1800..1829
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1836..1865
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2847..2865)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2848..2865)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2883..2899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2883..2899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2896..2918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2907..2923
                     /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(2931..2937,2938..2955,2956..2961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2976..2997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3114..3131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3285..3873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3365..3384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4044..4835,4836..4904))
                     /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     4667..4686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4905..5009)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5087..5542)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5224..5245)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5436..5455
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5668..5690
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5682..5699
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5683..5699
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5706..5722,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5706..5722,1..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 tatggtatca aagggcgaag
     1081 cagtaatcaa ggagttcatg cggtttaagg tacacatgga gggttccatg aatggccatg
     1141 aatttgagat cgaaggagag ggggaagggc gtccttacga aggaacacaa acagctaaac
     1201 ttaaggtaac taagggggga cccctcccct tttcctggga catactgtca ccacaattca
     1261 tgtacggcag ccgagccttt ataaaacatc cagctgatat acccgattat tataagcaaa
     1321 gttttcccga gggctttaag tgggaaaggg tcatgaactt cgaggacggg ggtgcggtaa
     1381 ctgtaaccca agacacctct ctcgaagacg ggacgcttat ttataaggtg aaactccggg
     1441 ggaccaattt tccccctgac ggacctgtca tgcagaagaa gacaatgggc tgggaggcgt
     1501 ccacagagcg cctgtatccg gaggatggcg tacttaaggg tgacataaaa atggcacttc
     1561 gtttgaagga cggtggccgt tatctcgctg actttaagac aacttataag gcgaaaaagc
     1621 cggtacagat gccaggagca tacaatgttg accgcaaatt ggacatcacc agccacaacg
     1681 aagactatac cgtcgttgaa cagtacgaac ggagtgaagg gaggcacagt acaggcggga
     1741 tggacgaatt gtataaagga tccgagcaga agctgataag cgaggaagac ctgggtagtg
     1801 aacagaagct gatctcagaa gaggaccttg gctcagaaca aaaactcatc tcagaggaag
     1861 atcttggatc aggatctgga tcaggatcgg gtagttaaga gctccgctgc gcttaaatgt
     1921 ctttcattat aatcaatgta taacctttat gtagtatttc aatctatgcc gctgtgtacg
     1981 tgcactgcgg tgcctatcct tcggcattag agagtcactg tttgtgtaga tcgtagctgc
     2041 atgtctgaca tctggttcat tagtgccgtt ttgagtctgc actgttgtgc aacttgatat
     2101 tcatcactgc acaccaaccg tgtgtttcct cctttgggtg caccctttaa tccttgtctt
     2161 ctcctttttt gtctctttcc ccctcgaaaa ggtgtcaaac tactgccgca taaactacgg
     2221 ttatcccaaa tttaagagaa agcaataaag catcagctag catgcctttg tctcaagaag
     2281 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     2341 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     2401 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     2461 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     2521 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     2581 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     2641 tgtgggaggg ctaactcgag tgcccattta gttggctttt cccttgtctc gtgtcttttc
     2701 cgtggaaagg ttcccggagt aatctgatgg cacagcaggg aggtgcgcct gcaggttggt
     2761 taggaagggg ggatgatgta aaagaagaaa atggggggat tcgagccccg cacaggtctc
     2821 tcaaattgga tgcatagctt gagtattcta tagtgtcacc taaatagctt ggcgtaatca
     2881 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2941 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3001 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3061 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3721 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3961 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4021 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4081 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aattacgata
     4141 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4201 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4261 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4321 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4381 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4441 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4501 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4561 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4621 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4681 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4741 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4801 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4861 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4921 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4981 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgatgcg
     5041 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     5101 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5161 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5221 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5281 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5341 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5401 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5461 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcactt
     5521 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     5581 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5641 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     5701 aattgtaata cgactcacta ta
//
