LOCUS       sequence-391376-        4224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for endogenously tagging Giardia genes with a triple Myc tag 
            (Puromycin selection).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4224)
  AUTHORS   Gourguechon S, Cande WZ
  TITLE     Rapid tagging and integration of genes in Giardia intestinalis.
  JOURNAL   Eukaryot Cell. 2011 Jan;10(1):142-5. doi: 10.1128/EC.00190-10. Epub 
            2010 Nov 29.
  PUBMED    21115739
REFERENCE   2  (bases 1 to 4224)
  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"; doi: 
            "10.1128/EC.00190-10"; journalName: "Eukaryot Cell"; date: 
            "2011-01"; volume: "10"; issue: "1"; pages: "142-5"
COMMENT     Sequence Label: pKS_3MYC_PAC
FEATURES             Location/Qualifiers
     source          1..4224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(101..120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     311..332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     595..617
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     609..626
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     610..626
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     684..700
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     684..700
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(734..750)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(735..751)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             768..797
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             810..839
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             852..881
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1303..1902
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1802..1822
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     promoter        complement(2046..2064)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2085..2101)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2085..2101)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2098..2120)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2109..2125
                     /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(2133..2139,2140..2157,2158..2163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2178..2199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2316..2333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2487..3075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2567..2586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3246..4037,4038..4106))
                     /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     3869..3888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4107..4211)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
       61 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      421 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat
      481 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      541 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      601 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
      661 agctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg
      721 atatcaagct tatcgatacc gtcgacctcg agcttaagag atccgaggag cagaagctga
      781 tctcggagga ggatctgctg cgtagtgaag aacaaaaact tataagtgaa gaagatttat
      841 tgaggtcaga ggaacagaaa ttaatttcag aagaggacct tttatgagga tcctgaatcg
      901 aattctttcc gtcgaccttt gcatcatcgt tgctcggtcg tctgcctcgt ttacgacctg
      961 cttgttcctg ggcctcaaat ggttagtggt tttccttctt ccaacggcgt tggtccatcc
     1021 tcggccgcgt gatcaaatgc cttcgagcgt atgtctgacc tgtttgtgaa tgacgatccg
     1081 tggctttact cagattcctg tgtttgtgaa taagaaatcc gtgaatttgt tggggtggaa
     1141 cggcgaagtg gcctctgcta caataagaac tggagagtgt gttcagaggc gaatgcgcaa
     1201 cgaactttgt cgttacacga cgtgtgcgcc tgtggtcttc gtgcggtttc cgcgcggaat
     1261 tttgcgcctt ttctaaaaaa ctaaccctaa aacaagaaaa tcatgggcac cgagtacaag
     1321 cccacggtgc gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc
     1381 gcgttcgccg actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg
     1441 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     1501 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     1561 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     1621 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     1681 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     1741 cccggagtgg aggcggccga gcgcgccggg gtgcctgcct tcctggagac ctccgcgccc
     1801 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     1861 gaaggaccgc gcacctggtg catgacccgc aagcccggtg ccctcgagtg agggctgaag
     1921 tgaatattta ccttttccgg gttcgacggg cgtgcttgac gatgcggaag tcgctgtccg
     1981 ttccaacggc tatctgcccc aggaccccca tcccggcgcc gatcagctgg taccagcttt
     2041 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     2101 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2161 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2221 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2281 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2341 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2401 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2461 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2521 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2581 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2641 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2701 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2761 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2821 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2881 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2941 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3001 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3061 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3121 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3181 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3241 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3301 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3361 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3421 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3481 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3541 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3601 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3661 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3721 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3781 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3841 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3901 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3961 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4021 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4081 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4141 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4201 ggggttccgc gcacatttcc ccga
//
