LOCUS       40924_20801        5865 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pFZZ-neo3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5865)
  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 5865)
  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 5865)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5865)
  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..5865
                     /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             4033..4812
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="DGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFV
                     KTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLA
                     PAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAP
                     AELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATR
                     DIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5180..5196)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5222..5240)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5247..5263)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5405..5860
                     /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 tcgatagaaa aatatttaaa aataaaaaat
     3181 aataagggtt ttgaataact cctttaattt aaatacacat ttttaaattt tttttagctc
     3241 tttaaatatt cataaaaata aaaaataact aactaaaaat aaataaaaag ataataatga
     3301 ttaaaggtat aatactgaat aagaaaaaac ataatagagt acttattttt tatatcacta
     3361 tttttaatat cttgaaagca aaactttttt atatatctta aaatatattg tatcgtttat
     3421 tcaattattt tctttaaatt tcaaatatat tgataaaaaa gatgacatgt tttttaaaga
     3481 aaacatgaaa tataaaatag ataaatatca attattttat ttattaaata tataagctgc
     3541 tcaaaacata gctcattcat caattataat atgtgaatca ttaattttca aaatattact
     3601 cattatttag gctatcattt attttttatt ttcaattatc cgtttctatt atattttaat
     3661 attaagttgt gattcttgaa ttttgtgtca tgaattattt gtaaatcttt ttatttctga
     3721 taaaaaatat aaattgattg actcatgatt taaatcatga gtcaacctaa ctaattttca
     3781 aaattcttct attctaaaat atagatgtga ttcttgaatc tctcttgaat ataaagtaat
     3841 tttttatatt tctgatataa ttcttagcta cgtgattcac gatttatgca atgatccata
     3901 taaaataatg taaatagtgt atatatatat attcgtcttt tttattcttt atataattta
     3961 aaaaaattaa aaaaatttaa taaagctcta ataaaataaa taataatact aaacttaaaa
     4021 taatgaagct tggatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4081 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4141 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4201 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4261 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4321 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4381 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4441 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4501 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4561 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4621 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4681 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4741 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4801 gacgagttct tctgagatcc ttaaattaaa aattcaatat atattttaca acttcatata
     4861 aataaatata ttatataaaa ttaattttta gtgtattaat attaacatta aagcaccaaa
     4921 aaaacgtgtt aatatactac tataaaatat aatttattcc aaattgacta aaatcattat
     4981 tttacaactc atttgtatat atattttatg tcaattattt tttttaactt tctaaaaaaa
     5041 aaaattcctc ttcacataca tgttagctct taaaaatttg tctgcaaatc caataataat
     5101 attttttttt gccattaaat tttcaaattt ttactggaaa aatgcagccc gggggatctg
     5161 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg
     5221 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     5281 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     5341 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     5401 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5461 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5521 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5581 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5641 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5701 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5761 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5821 tttaacgcga attttaacaa aatattaacg cttacaattt aggtg
//
