LOCUS       40924_7751        5127 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Giardia integration vector pc-CycB-3HA-PAC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  AUTHORS   Gourguechon S, Cande WZ.
  TITLE     Rapid tagging and integration of genes in Giardia intestinalis
  JOURNAL   Eukaryotic Cell (2010) In press
  PUBMED    21115739
REFERENCE   2  (bases 1 to 5127)
  AUTHORS   Gourguechon S, Cande WZ.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-NOV-2010) Mol and Cell Biology, University of 
            California, Berkeley, 345 LSA, Berkeley, CA 94720-3200, USA
REFERENCE   3  (bases 1 to 5127)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5127)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Eukaryotic 
            Cell (2010) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-NOV-2010) Mol and Cell Biology, University of California, 
            Berkeley, 345 LSA, Berkeley, CA 94720-3200, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1687..1713
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             1717..1743
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             1747..1773
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             2195..2788
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     promoter        complement(2939..2957)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2978..2994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3002..3018)
                     /label=lac operator
                     /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(3026..3056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3071..3092)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3380..3968)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4142..4999)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5000..5104)
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccat
      661 tccacaattg acggcaatcg ctacggctat agcccgaggg gcgcctctaa cgatcgttta
      721 cctgagcgcc gacgctcgaa aaggggtgag gattatcgga aagaacacac ttacaaaccc
      781 accatttcca agtttgcgca ggcgctgcct cgctcagatg ttccgtgtca cgaacggctg
      841 tattcagcca gacctagtgt tcaacggaac aagaatgcag atggtgagga ctacgaggca
      901 ccggcactct ctccgcgttt gcaggagttt gtgaacaaca gcagttttga gttgttccgt
      961 ccccacatat caaagcgtgc aagggcttta gaggtcgaag cgcctttcta cgatcgtctc
     1021 catccatcca aggcctctaa cagacttgtt ggcatttgct gtcttttcat tgcagcaaag
     1081 tacgagcaaa gctatattcc ttcacttagg cattttgtgg aggcgctgac ccaggtcgac
     1141 ccggaattcc tcgtcaattg taagtgcgag ggtaaccgtt cgggaccgtg ctacaggtgc
     1201 agtcgcgaag ctataataga tacggaagaa tctatcctgc atacgctgga tttccagctg
     1261 gctatgccca caattctcac attcctacga aggtatgcca gaatttcgga aatgaagaat
     1321 cgagaccgtt acattgcatt ttatatctct gaactctcgt gtctgacgac aagtatcttc
     1381 aaatttaagc ctagcgagat tgcggcaggc tgcattgcga tgtcccgtag attgactcgg
     1441 aagccgactt gggacgaaac cctcgaaaag tattctggat acactgagag cgatacaaaa
     1501 aaggttatgg cggagatttc tacgatcatg aagaaatacg cttgtaaccc gaaggccaga
     1561 tacataaggc gcaaatactc gcaagaggat cgcttctacg gggtgtcctt gatgattgag
     1621 actgtggtaa cgcaggcgct aactcttaat acaaaggaag caaagcccgg gctcgagctt
     1681 aaggaatatc cttatgacgt cccagactac gcggagtacc catacgatgt acctgattat
     1741 gctgagtacc catacgacgt accagattac gctggatcct gaatcgaatt ctttccgtcg
     1801 acctttgcat catcgttgct cggtcgtctg cctcgtttac gacctgcttg ttcctgggcc
     1861 tcaaatggtt agtggttttc cttcttccaa cggcgttggt ccatcctcgg ccgcgtgatc
     1921 aaatgccttc gagcgtatgt ctgacctgtt tgtgaatgac gatccgtggc tttactcaga
     1981 ttcctgtgtt tgtgaataag aaatccgtga atttgttggg gtggaacggc gaagtggcct
     2041 ctgctacaat aagaactgga gagtgtgttc agaggcgaat gcgcaacgaa ctttgtcgtt
     2101 acacgacgtg tgcgcctgtg gtcttcgtgc ggtttccgcg cggaattttg cgccttttct
     2161 aaaaaactaa ccctaaaaca agaaaatcat gggcaccgag tacaagccca cggtgcgcct
     2221 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     2281 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     2341 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     2401 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     2461 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     2521 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     2581 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     2641 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     2701 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag tgcccgaagg accgcgcgac
     2761 ctggtgcatg acccgcaagc ccggtgccct cgagtgaggg ctgaagtgaa tatttacctt
     2821 ttccgggttc gacgggcgtg cttgacgatg cggaagtcgc tgtccgttcc aacggctatc
     2881 tgccccagga cccccatccc ggcgccgatc agctggtacc gagctccagc ttttgttccc
     2941 tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3001 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3061 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3121 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3181 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3241 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3301 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3361 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3421 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3481 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3541 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3601 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3661 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3721 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3781 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3841 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3901 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3961 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4021 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4081 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4141 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4201 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4261 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4321 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4381 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4441 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4501 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4561 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4621 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4681 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4741 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4801 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4861 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4921 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4981 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5041 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5101 cgcgcacatt tccccgaaaa gtgccac
//
