LOCUS       40924_24049        5418 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pHA-neo3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  AUTHORS   Kataoka K, Schoeberl UE, Mochizuki K.
  TITLE     Modules for C-terminal epitope tagging of Tetrahymena genes
  JOURNAL   J. Microbiol. Methods 82 (3), 342-346 (2010)
  PUBMED    20624430
REFERENCE   2  (bases 1 to 5418)
  AUTHORS   Kataoka K, Mochizuki K.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2010) Contact:Kazufumi Mochizuki Institute of 
            Molecular Biotechnology of the Austrian Academy of Sciences (IMBA); 
            Dr. Bohr-Gasse 3, Vienna A-1030, Austria
REFERENCE   3  (bases 1 to 5418)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5418)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2010"; volume: "82"; issue: "3"; pages: 
            "342-346"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUN-2010) Contact:Kazufumi Mochizuki Institute of Molecular 
            Biotechnology of the Austrian Academy of Sciences (IMBA); Dr. 
            Bohr-Gasse 3, Vienna A-1030, Austria"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /label=AmpR promoter
     CDS             126..983
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1157..1745
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2069..2099
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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)."
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2247..2273
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_feature    2280..2678
                     /label=BTU1 3'-flanking
                     /note="BTU1 3'-flanking"
     CDS             3586..4365
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="DGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFV
                     KTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLA
                     PAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAP
                     AELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATR
                     DIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4733..4749)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4775..4793)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4800..4816)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4958..5413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctcgaaat taaccctcac taaagggaac aaaagctgga gctccaccgc ggtggcggcc
     2221 gctctagagc atgcgctagc ggatcctatc cttatgatgt tcctgattat gctactagtt
     2281 gagcgaactg aatcggtcag ctaaaccaac caatcaacat aataaacttt attattttta
     2341 cttaagcatc ttactgttgt tgtaatagta gagaaagaaa tacccaatta acttcattca
     2401 cataacatta atatctataa acatcttttt tctcacatat atacaactct ctaaatcaac
     2461 aaataacttt ttaaaaataa tggatatata ttaacaaaat aatatatctc tttttacaaa
     2521 atagttctta tataaatacg tattctgcac tcacccgcat ttttcacaac aaaaacatac
     2581 caaaaaaatt cttacttcta catgtttcct ttcttattat tacaaaatta ttttataaat
     2641 agcatacaaa aataaataca ataaaaaaat aaacaaaatc cagatccccc gggctgcagg
     2701 aattcgatag aaaaatattt aaaaataaaa aataataagg gttttgaata actcctttaa
     2761 tttaaataca catttttaaa ttttttttag ctctttaaat attcataaaa ataaaaaata
     2821 actaactaaa aataaataaa aagataataa tgattaaagg tataatactg aataagaaaa
     2881 aacataatag agtacttatt ttttatatca ctatttttaa tatcttgaaa gcaaaacttt
     2941 tttatatatc ttaaaatata ttgtatcgtt tattcaatta ttttctttaa atttcaaata
     3001 tattgataaa aaagatgaca tgttttttaa agaaaacatg aaatataaaa tagataaata
     3061 tcaattattt tatttattaa atatataagc tgctcaaaac atagctcatt catcaattat
     3121 aatatgtgaa tcattaattt tcaaaatatt actcattatt taggctatca tttatttttt
     3181 attttcaatt atccgtttct attatatttt aatattaagt tgtgattctt gaattttgtg
     3241 tcatgaatta tttgtaaatc tttttatttc tgataaaaaa tataaattga ttgactcatg
     3301 atttaaatca tgagtcaacc taactaattt tcaaaattct tctattctaa aatatagatg
     3361 tgattcttga atctctcttg aatataaagt aattttttat atttctgata taattcttag
     3421 ctacgtgatt cacgatttat gcaatgatcc atataaaata atgtaaatag tgtatatata
     3481 tatattcgtc ttttttattc tttatataat ttaaaaaaat taaaaaaatt taataaagct
     3541 ctaataaaat aaataataat actaaactta aaataatgaa gcttggatgg attgcacgca
     3601 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3661 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3721 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3781 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3841 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3901 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3961 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4021 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4081 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4141 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4201 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4261 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4321 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaga tccttaaatt
     4381 aaaaattcaa tatatatttt acaacttcat ataaataaat atattatata aaattaattt
     4441 ttagtgtatt aatattaaca ttaaagcacc aaaaaaacgt gttaatatac tactataaaa
     4501 tataatttat tccaaattga ctaaaatcat tattttacaa ctcatttgta tatatatttt
     4561 atgtcaatta ttttttttaa ctttctaaaa aaaaaaattc ctcttcacat acatgttagc
     4621 tcttaaaaat ttgtctgcaa atccaataat aatatttttt tttgccatta aattttcaaa
     4681 tttttactgg aaaaatgcag cccgggggat ctgaattcga tatcaagctt atcgataccg
     4741 tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacaattca
     4801 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4861 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4921 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     4981 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5041 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5101 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5161 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5221 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5281 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5341 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5401 acgcttacaa tttaggtg
//
