LOCUS       40924_24054        5435 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pHA-neo4 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5435)
  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 5435)
  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 5435)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5435)
  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..5435
                     /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             3590..4381
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4750..4766)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4792..4810)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4817..4833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4975..5430
                     /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 acaatttatt tctaaaaaat atttaaaaat aaaaaataat aagggttttg
     2761 aataactcct ttaatttaaa tacacatttt taaatttttt ttagctcttt aaatattcat
     2821 aaaaataaaa aataactaac taaaaataaa taaaaagata ataatgatta aaggtataat
     2881 actgaataag aaaaaacata atagagtact tattttttat atcactattt ttaatatctt
     2941 gaaagcaaaa cttttttata tatcttaaaa tatattgtat cgtttattca attattttct
     3001 ttaaatttca aatatattga taaaaaagat gacatgtttt ttaaagaaaa catgaaatat
     3061 aaaatagata aatatcaatt attttattta ttaaatatat aagctgctca aaacatagct
     3121 cattcatcaa ttataatatg tgaatcatta attttcaaaa tattactcat tatttaggct
     3181 atcatttatt ttttattttc aattatccgt ttctattata ttttaatatt aagttgtgat
     3241 tcttgaattt tgtgtcatga attatttgta aatcttttta tttctgataa aaaatataaa
     3301 ttgattgact catgatttaa atcatgagtc aacctaacta attttcaaaa ttcttctatt
     3361 ctaaaatata gatgtgattc ttgaatctct cttgaatata aagtaatttt ttatatttct
     3421 gatataattc ttagctacgt gattcacgat ttatacaatg atccatataa aataatgtaa
     3481 atagtgtata tatatatatt cgtctttttt attctttata taatttaaaa aaattaaaaa
     3541 aatttaataa agctctaata aaataaataa taatactaaa cttaaaataa tgattgaaca
     3601 agatggttta cacgctggtt ctcccgccgc ttgggtcgaa agacttttcg gttatgactg
     3661 ggctcaacaa accatcggtt gctctgatgc cgccgtcttc cgtctttctg ctcaaggtcg
     3721 tcctgttctt ttcgtcaaga ccgacctttc tggtgccctt aatgaacttc aagatgaagc
     3781 tgcccgtctt tcttggcttg ccaccaccgg tgttccttgc gctgctgtcc ttgacgttgt
     3841 cactgaagcc ggtagagact ggcttctttt aggtgaagtc cccggtcaag atcttctttc
     3901 ttctcacctt gctcctgccg aaaaagtttc tatcatggct gatgctatgc gtcgtcttca
     3961 tacccttgat cccgctacct gccctttcga ccaccaagcc aaacatcgta tcgaacgtgc
     4021 tcgtactcgt atggaagccg gtcttgtcga tcaagatgat cttgacgaag aacatcaagg
     4081 tcttgcccct gccgaacttt tcgccagact taaggcccgt atgcccgacg gtgaagatct
     4141 tgtcgtcacc catggtgatg cctgcttacc caatatcatg gttgaaaatg gtcgtttttc
     4201 tggtttcatc gactgtggtc gtcttggtgt cgccgaccgt tatcaagata ttgccttagc
     4261 tacccgtgat attgctgaag aacttggtgg tgaatgggct gaccgtttcc ttgtccttta
     4321 cggtatcgcc gctcccgatt ctcaacgtat cgccttctat cgtcttcttg acgaattctt
     4381 ctgagatcct taaattaaaa attcaatata tatttacaaa ctttcatata aaataaatat
     4441 attatataaa attaattttt agtgtattat attaacatta aagcaccaaa aaaacgtgtt
     4501 aatatactac tataaaatat aatttattcc aaattgacta aaatcattat tttacaactc
     4561 atttgtatat atattttatg tcaattattt tttttaactt tctaaaaaaa aaaattcctc
     4621 ttcacataca tgttagctct taaaaatttg tctgcaaatc caataataat attttttttt
     4681 gccattaaat tttcaaattt ttactggaaa aatgcagccc gggggatctg aattcgatat
     4741 caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg ccctatagtg
     4801 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4861 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4921 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     4981 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5041 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5101 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5161 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5221 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5281 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5341 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5401 attttaacaa aatattaacg cttacaattt aggtg
//
