LOCUS       sequence-355617-        5146 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 5146)
  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 5146)
  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: pPOTv7-blast-10HA
FEATURES             Location/Qualifiers
     source          1..5146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             960..986
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             993..1019
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1026..1052
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1059..1085
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1092..1118
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1125..1151
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1158..1184
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1191..1217
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1224..1250
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1257..1283
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2271..2289)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2272..2289)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2307..2323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2307..2323)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2320..2342)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2331..2347
                     /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(2355..2361,2362..2379,2380..2385))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2400..2421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2538..2555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2709..3297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2789..2808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3468..4259,4260..4328))
                     /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     4091..4110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4329..4433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4511..4966)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4648..4669)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4860..4879
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5092..5114
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5106..5123
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5107..5123
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5130..5146,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5130..5146,1..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 tctaagcttt
      961 atccttatga tgtgccagat tacgctggtt cctatcccta tgacgttcct gactacgctg
     1021 ggtcgtaccc ttatgatgtg ccggattacg ccggaagcta cccatacgat gtaccggact
     1081 acgcagggtc atatccttac gatgttccag actatgctgg ctcctatccg tacgatgtcc
     1141 ccgactatgc aggatcatac ccgtatgacg ttccagatta tgccgggtcg tacccttatg
     1201 acgtcccaga ttatgccggt tcttatccgt atgatgtgcc ggactacgcg gggtcctatc
     1261 cgtatgatgt acccgattac gcgggatccg gatcaggatc tggatcagga tcgggtagtt
     1321 aagagctccg ctgcgcttaa atgtctttca ttataatcaa tgtataacct ttatgtagta
     1381 tttcaatcta tgccgctgtg tacgtgcact gcggtgccta tccttcggca ttagagagtc
     1441 actgtttgtg tagatcgtag ctgcatgtct gacatctggt tcattagtgc cgttttgagt
     1501 ctgcactgtt gtgcaacttg atattcatca ctgcacacca accgtgtgtt tcctcctttg
     1561 ggtgcaccct ttaatccttg tcttctcctt ttttgtctct ttccccctcg aaaaggtgtc
     1621 aaactactgc cgcataaact acggttatcc caaatttaag agaaagcaat aaagcatcag
     1681 ctagcatgcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca
     1741 acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc
     1801 gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg
     1861 tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa
     1921 atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc
     1981 atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc
     2041 gtgaattgct gccctctggt tatgtgtggg agggctaact cgagtgccca tttagttggc
     2101 ttttcccttg tctcgtgtct tttccgtgga aaggttcccg gagtaatctg atggcacagc
     2161 agggaggtgc gcctgcaggt tggttaggaa ggggggatga tgtaaaagaa gaaaatgggg
     2221 ggattcgagc cccgcacagg tctctcaaat tggatgcata gcttgagtat tctatagtgt
     2281 cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2341 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2401 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2461 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2521 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2581 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2641 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2701 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2761 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2821 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2881 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2941 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3001 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3061 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3121 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3241 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3301 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3361 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3421 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3541 cccgtcgtgt agataattac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3601 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3661 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3721 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3781 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3841 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3901 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3961 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4021 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4081 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4141 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4201 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4261 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4321 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4381 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4441 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4501 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4561 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4621 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4681 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4741 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4801 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4861 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4921 gtaaccacca cacccgccgc acttaatgcg ccgctacagg gcgcgtccat tcgccattca
     4981 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     5041 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     5101 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actata
//
