LOCUS       40924_9086        5007 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Giardia integration vector pcAUK-3HA-NEO, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5007)
  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 5007)
  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 5007)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5007)
  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..5007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /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"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1390..1416
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             1420..1446
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             1450..1476
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             1880..2671
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(2819..2837)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2858..2874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2882..2898)
                     /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(2906..2936)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2951..2972)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3260..3848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4022..4879)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4880..4984)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatgagtc
      721 tttttggtcc tcgaagtggc tccgtacggc gagctttacc aaaggttaca gcagttcggg
      781 aagtttcctc tacccgttgt aagcaagatt attcgagacg tggcacaggc gatacagtat
      841 cttcacagaa agaacatctt ccatagggac cttaaggcag aaaacatctt gatttgcaag
      901 gggaaagaga ccaaagagca cacagatgcg cacaattcag atgactccat ttctgtgcat
      961 gaacacgagc tagtccgaat ggcacactac acatacaaga ttgcagactt tggatggtcc
     1021 gtgcaccatc caacccacgg tggccgacga cgcacccagt gcggaactct cgactacctt
     1081 cccccggagg tcatgctcgg acaatcttat gataaggctt gcgatatctg gtcactcgga
     1141 gccctgtgct atgagctcat ctgtggcacc gcgcccttct accacgacga aatcaagatc
     1201 acacgacaaa atatagccaa cgtagagtat agctttacta aggacttcag cccagccagc
     1261 aaagatttca tccagcggat gttaattaga agcccagagg cccggatctc catagaggat
     1321 atcctccggc atccattcct gagacagaca gatcacagga gtaaggtccc caagctcgag
     1381 cttaaggaat atccttatga cgtcccagac tacgcggagt acccatacga tgtacctgat
     1441 tatgctgagt acccatacga cgtaccagat tacgctggat cctgaatcga attctttgca
     1501 tcatcgttgc tcggtcgtct gcctcgttta cgacctgctt gttcctgggc ctcaaatggt
     1561 tagtggtttt ccttcttcca acggcgttgg tccatcctcg gccgcgtgat caaatgcctt
     1621 cgagcgtatg tctgacctgt ttgtgaatga cgatccgtgg ctttactcag attcctgtgt
     1681 ttgtgaataa gaaatccgtg aatttgttgg ggtggaacgg cgaagtggcc tctgctacaa
     1741 taagaactgg ggagtgtgtt cagaggcgaa tgcgcaacga actttgtcgt tacacgacgt
     1801 gtgcgcctgt ggtcttcgtg cggtttccgc gcggaatttt gcgccttttc taaaaaacta
     1861 accctaaaac aagaaaatca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     1921 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     1981 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     2041 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     2101 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     2161 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     2221 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     2281 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     2341 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     2401 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     2461 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     2521 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     2581 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     2641 cgccttctat cgccttcttg acgagttctt ctgagtcgag tgagggctga agtgaatatt
     2701 taccttttcc gggttcgacg ggcgtgcttg acgatgcgga agtcgctgtc cgttccaacg
     2761 gctatctgcc ccaggacccc catcccggcg ccgatcagct ggtacccagc ttttgttccc
     2821 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2881 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2941 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3001 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3061 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3661 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     3721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4981 cgcgcacatt tccccgaaaa gtgccac
//
