LOCUS       40924_20806        5882 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pFZZ-neo4 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5882)
  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 5882)
  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 5882)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5882)
  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..5882
                     /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..2312
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2316..2336
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             2361..2534
                     /codon_start=1
                     /label=ProtA
                     /note="IgG-binding unit of Staphylococcus aureus protein A"
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     misc_feature    2727..3125
                     /label=BTU1 3' flanking
                     /note="BTU1 3' flanking"
     CDS             4037..4828
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5197..5213)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5239..5257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5264..5280)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5422..5877
                     /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 ggatccgatt acaaagatca tgatggtgat tacaaagatc
     2281 acgatattga ttacaaagat gatgatgata aagctgaaaa tctttatttt caaggtgctt
     2341 ctgctcaaca tgatgaagct gttgataaca aatttaataa ggaacaacaa aatgcttttt
     2401 acgaaatttt acatcttcct aatttaaatg aagaacaaag aaatgctttc attcaatcat
     2461 taaaagatga tccttcacaa tctgctaatt tattagctga agctaaaaaa ttaaatgatg
     2521 ctcaagctcc taaagttgat aataaattca acaaagaaca acaaaacgct ttttatgaaa
     2581 ttcttcattt acctaacctt aatgaagaac aaagaaacgc tttcattcaa tctttaaagg
     2641 atgatccttc tcaatctgct aatttacttg ctgaagctaa aaagttaaat ggtgctcaag
     2701 ctcctaaggt tgatgctaat actagttgag cgaactgaat cggtcagcta aaccaaccaa
     2761 tcaacataat aaactttatt atttttactt aagcatctta ctgttgttgt aatagtagag
     2821 aaagaaatac ccaattaact tcattcacat aacattaata tctataaaca tcttttttct
     2881 cacatatata caactctcta aatcaacaaa taacttttta aaaataatgg atatatatta
     2941 acaaaataat atatctcttt ttacaaaata gttcttatat aaatacgtat tctgcactca
     3001 cccgcatttt tcacaacaaa aacataccaa aaaaattctt acttctacat gtttcctttc
     3061 ttattattac aaaattattt tataaatagc atacaaaaat aaatacaata aaaaaataaa
     3121 caaaatccag atcccccggg ctgcaggaat tcgatagaca atttatttct aaaaaatatt
     3181 taaaaataaa aaataataag ggttttgaat aactccttta atttaaatac acatttttaa
     3241 atttttttta gctctttaaa tattcataaa aataaaaaat aactaactaa aaataaataa
     3301 aaagataata atgattaaag gtataatact gaataagaaa aaacataata gagtacttat
     3361 tttttatatc actattttta atatcttgaa agcaaaactt ttttatatat cttaaaatat
     3421 attgtatcgt ttattcaatt attttcttta aatttcaaat atattgataa aaaagatgac
     3481 atgtttttta aagaaaacat gaaatataaa atagataaat atcaattatt ttatttatta
     3541 aatatataag ctgctcaaaa catagctcat tcatcaatta taatatgtga atcattaatt
     3601 ttcaaaatat tactcattat ttaggctatc atttattttt tattttcaat tatccgtttc
     3661 tattatattt taatattaag ttgtgattct tgaattttgt gtcatgaatt atttgtaaat
     3721 ctttttattt ctgataaaaa atataaattg attgactcat gatttaaatc atgagtcaac
     3781 ctaactaatt ttcaaaattc ttctattcta aaatatagat gtgattcttg aatctctctt
     3841 gaatataaag taatttttta tatttctgat ataattctta gctacgtgat tcacgattta
     3901 tacaatgatc catataaaat aatgtaaata gtgtatatat atatattcgt cttttttatt
     3961 ctttatataa tttaaaaaaa ttaaaaaaat ttaataaagc tctaataaaa taaataataa
     4021 tactaaactt aaaataatga ttgaacaaga tggtttacac gctggttctc ccgccgcttg
     4081 ggtcgaaaga cttttcggtt atgactgggc tcaacaaacc atcggttgct ctgatgccgc
     4141 cgtcttccgt ctttctgctc aaggtcgtcc tgttcttttc gtcaagaccg acctttctgg
     4201 tgcccttaat gaacttcaag atgaagctgc ccgtctttct tggcttgcca ccaccggtgt
     4261 tccttgcgct gctgtccttg acgttgtcac tgaagccggt agagactggc ttcttttagg
     4321 tgaagtcccc ggtcaagatc ttctttcttc tcaccttgct cctgccgaaa aagtttctat
     4381 catggctgat gctatgcgtc gtcttcatac ccttgatccc gctacctgcc ctttcgacca
     4441 ccaagccaaa catcgtatcg aacgtgctcg tactcgtatg gaagccggtc ttgtcgatca
     4501 agatgatctt gacgaagaac atcaaggtct tgcccctgcc gaacttttcg ccagacttaa
     4561 ggcccgtatg cccgacggtg aagatcttgt cgtcacccat ggtgatgcct gcttacccaa
     4621 tatcatggtt gaaaatggtc gtttttctgg tttcatcgac tgtggtcgtc ttggtgtcgc
     4681 cgaccgttat caagatattg ccttagctac ccgtgatatt gctgaagaac ttggtggtga
     4741 atgggctgac cgtttccttg tcctttacgg tatcgccgct cccgattctc aacgtatcgc
     4801 cttctatcgt cttcttgacg aattcttctg agatccttaa attaaaaatt caatatatat
     4861 ttacaaactt tcatataaaa taaatatatt atataaaatt aatttttagt gtattatatt
     4921 aacattaaag caccaaaaaa acgtgttaat atactactat aaaatataat ttattccaaa
     4981 ttgactaaaa tcattatttt acaactcatt tgtatatata ttttatgtca attatttttt
     5041 ttaactttct aaaaaaaaaa attcctcttc acatacatgt tagctcttaa aaatttgtct
     5101 gcaaatccaa taataatatt tttttttgcc attaaatttt caaattttta ctggaaaaat
     5161 gcagcccggg ggatctgaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg
     5221 gcccggtacc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     5281 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     5341 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     5401 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5461 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5521 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5581 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5641 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5701 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5761 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5821 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     5881 tg
//
