LOCUS       sequence-391375-        4368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for endogenously tagging Giardia genes with a triple HA tag 
            (G418 selection).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4368)
  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 4368)
  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_3HA_NEO
FEATURES             Location/Qualifiers
     source          1..4368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(106..125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     316..337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     600..622
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     614..631
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     615..631
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     689..705
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     689..705
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(739..755)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(740..756)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             767..793
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             797..823
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             827..853
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1257..2051
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1311..1330)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1921..1940
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(2195..2213)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2234..2250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2234..2250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2247..2269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2258..2274
                     /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(2282..2288,2289..2306,2307..2312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2327..2348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2465..2482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2636..3224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2716..2735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3395..4186,4187..4255))
                     /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     4018..4037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4256..4360)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
       61 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
      121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
      181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
      241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
      301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
      361 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
      421 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
      481 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      541 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      601 ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga
      661 attggagctc caccgcggtg gcggccgctc tagaactagt ggatcccccg ggctgcagga
      721 attcgatatc aagcttatcg ataccgtcga cctcgagctt aaggaatatc cttatgacgt
      781 cccagactac gcggagtacc catacgatgt acctgattat gctgagtacc catacgacgt
      841 accagattac gctggatcct gaatcgaatt ctttgcatca tcgttgctcg gtcgtctgcc
      901 tcgtttacga cctgcttgtt cctgggcctc aaatggttag tggttttcct tcttccaacg
      961 gcgttggtcc atcctcggcc gcgtgatcaa atgccttcga gcgtatgtct gacctgtttg
     1021 tgaatgacga tccgtggctt tactcagatt cctgtgtttg tgaataagaa atccgtgaat
     1081 ttgttggggt ggaacggcga agtggcctct gctacaataa gaactgggga gtgtgttcag
     1141 aggcgaatgc gcaacgaact ttgtcgttac acgacgtgtg cgcctgtggt cttcgtgcgg
     1201 tttccgcgcg gaattttgcg ccttttctaa aaaactaacc ctaaaacaag aaaatcatga
     1261 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     1321 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     1381 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     1441 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     1501 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     1561 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     1621 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     1681 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     1741 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     1801 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     1861 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     1921 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     1981 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2041 agttcttctg agtcgagtga gggctgaagt gaatatttac cttttccggg ttcgacgggc
     2101 gtgcttgacg atgcggaagt cgctgtccgt tccaacggct atctgcccca ggacccccat
     2161 cccggcgccg atcagctggt accagctttt gttcccttta gtgagggtta atttcgagct
     2221 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2281 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2341 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2401 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2461 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2521 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2581 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2641 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2701 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2761 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2821 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2881 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2941 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3001 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3061 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3121 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3181 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3241 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3301 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3361 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3421 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3481 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3541 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3601 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3661 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3721 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3781 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3841 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3901 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3961 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4021 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4081 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4141 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4201 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4261 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4321 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttc
//
