LOCUS       Exported                7245 bp ds-DNA     circular SYN 20-AUG-2019
DEFINITION  Transformation vector pFZZ-neo3 DNA, complete sequence.
ACCESSION   AB570111
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7245)
  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 7245)
  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 7245)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Nov 27, 2019 from SnapGene 4.3.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7245
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..2240
                     /label=pBlueScript SK(+)
                     /note="pBlueScript SK(+)"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             126..986
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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"
     primer_bind     complement(2220..2267)
                     /label=Vector_MS1_Primer_R
     primer_bind     2227..2279
                     /label=Q22MS1_5'UTR_F
     misc_feature    2247..2666
                     /label=Q22MS1_5'UTR
     primer_bind     2601..2629
                     /label=Q22MS1_5'UTR_For
     primer_bind     complement(2626..2688)
                     /label=Q22MS1_5'UTR_R
     primer_bind     2641..2702
                     /label=Vector_MS1_FLAG_F
     misc_feature    2667..2732
                     /gene="FLAG"
                     /label=FLAG
     CDS             2667..2732
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2736..2756
                     /label=TEV protease recognition site
                     /note="TEV protease recognition site"
     CDS             2736..2756
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    2766..3140
                     /label=ZZ domain of protein A codon-optimized for Tetrahy
                     /note="ZZ domain of protein A codon-optimized for 
                     Tetrahymena"
     misc_feature    3147..3577
                     /label=BTU1 3' flanking
                     /note="BTU1 3' flanking"
     misc_feature    3578..4442
                     /label=MTT1_5'
     primer_bind     complement(4434..4465)
                     /label=M_Neo3R
     misc_feature    4443..5566
                     /label=neo3
                     /note="neo3"
     primer_bind     complement(5531..5583)
                     /label=Vector_MS1_Neo3_R
     primer_bind     5544..5592
                     /label=Q22MS1_3'UTR_F
     misc_feature    5567..6526
                     /label=Q22MS1_3'UTR
     primer_bind     complement(5601..5644)
                     /label=Q22MS1_3'UTR_Rev
     primer_bind     complement(6495..6546)
                     /label=Q22MS1_3'UTR_R
     primer_bind     6507..6554
                     /label=Vector_MS1_Primer_F
     misc_feature    6527..7245
                     /label=pBlueScript SK(+)(1)
                     /note="pBlueScript SK(+)"
     primer_bind     complement(6560..6576)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6602..6620)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6627..6643)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6785..7240
                     /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 ggatcctaaa ttttgtatca aaaatagaaa aattaaatct
     2281 ttttcaagat taaaatatat tttcttgtta tttatttaat aaaaattaaa atattatttt
     2341 atttattttt gaaaaataga atgtttttaa aacttattag aattaattat taataaaaat
     2401 tatttatttt taagtaaagt gaaataaaca atttatcagt tttacatttt tagggttatt
     2461 aacagttaac actgagatag gttgttttaa aaattttaat tatagtcttt gaacttagaa
     2521 attatttttt ttgaaaatca tttttttaaa aataaaaaat aattatttat taattaatta
     2581 ttgaatataa aaaagaagcg caaattcaac atttaaaata ttcagcaact taaaaaatta
     2641 tatatttttt taatttttaa atatatgatt acaaagatca tgatggtgat tacaaagatc
     2701 acgatattga ttacaaagat gatgatgata aagctgaaaa tctttatttt caaggtgctt
     2761 ctgctcaaca tgatgaagct gttgataaca aatttaataa ggaacaacaa aatgcttttt
     2821 acgaaatttt acatcttcct aatttaaatg aagaacaaag aaatgctttc attcaatcat
     2881 taaaagatga tccttcacaa tctgctaatt tattagctga agctaaaaaa ttaaatgatg
     2941 ctcaagctcc taaagttgat aataaattca acaaagaaca acaaaacgct ttttatgaaa
     3001 ttcttcattt acctaacctt aatgaagaac aaagaaacgc tttcattcaa tctttaaagg
     3061 atgatccttc tcaatctgct aatttacttg ctgaagctaa aaagttaaat ggtgctcaag
     3121 ctcctaaggt tgatgctaat actagttgag cgaactgaat cggtcagcta aaccaaccaa
     3181 tcaacataat aaactttatt atttttactt aagcatctta ctgttgttgt aatagtagag
     3241 aaagaaatac ccaattaact tcattcacat aacattaata tctataaaca tcttttttct
     3301 cacatatata caactctcta aatcaacaaa taacttttta aaaataatgg atatatatta
     3361 acaaaataat atatctcttt ttacaaaata gttcttatat aaatacgtat tctgcactca
     3421 cccgcatttt tcacaacaaa aacataccaa aaaaattctt acttctacat gtttcctttc
     3481 ttattattac aaaattattt tataaatagc atacaaaaat aaatacaata aaaaaataaa
     3541 caaaatccag atcccccggg ctgcaggaat tcgatagaaa aatatttaaa aataaaaaat
     3601 aataagggtt ttgaataact cctttaattt aaatacacat ttttaaattt tttttagctc
     3661 tttaaatatt cataaaaata aaaaataact aactaaaaat aaataaaaag ataataatga
     3721 ttaaaggtat aatactgaat aagaaaaaac ataatagagt acttattttt tatatcacta
     3781 tttttaatat cttgaaagca aaactttttt atatatctta aaatatattg tatcgtttat
     3841 tcaattattt tctttaaatt tcaaatatat tgataaaaaa gatgacatgt tttttaaaga
     3901 aaacatgaaa tataaaatag ataaatatca attattttat ttattaaata tataagctgc
     3961 tcaaaacata gctcattcat caattataat atgtgaatca ttaattttca aaatattact
     4021 cattatttag gctatcattt attttttatt ttcaattatc cgtttctatt atattttaat
     4081 attaagttgt gattcttgaa ttttgtgtca tgaattattt gtaaatcttt ttatttctga
     4141 taaaaaatat aaattgattg actcatgatt taaatcatga gtcaacctaa ctaattttca
     4201 aaattcttct attctaaaat atagatgtga ttcttgaatc tctcttgaat ataaagtaat
     4261 tttttatatt tctgatataa ttcttagcta cgtgattcac gatttatgca atgatccata
     4321 taaaataatg taaatagtgt atatatatat attcgtcttt tttattcttt atataattta
     4381 aaaaaattaa aaaaatttaa taaagctcta ataaaataaa taataatact aaacttaaaa
     4441 taatgaagct tggatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4501 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4561 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4621 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4681 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4741 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4801 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4861 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4921 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4981 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5041 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5101 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5161 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     5221 gacgagttct tctgagatcc ttaaattaaa aattcaatat atattttaca acttcatata
     5281 aataaatata ttatataaaa ttaattttta gtgtattaat attaacatta aagcaccaaa
     5341 aaaacgtgtt aatatactac tataaaatat aatttattcc aaattgacta aaatcattat
     5401 tttacaactc atttgtatat atattttatg tcaattattt tttttaactt tctaaaaaaa
     5461 aaaattcctc ttcacataca tgttagctct taaaaatttg tctgcaaatc caataataat
     5521 attttttttt gccattaaat tttcaaattt ttactggaaa aatgcaaatc tcttatagga
     5581 gtcaaatatc tgaaaatttt gaatttataa atcaattata attataacta aaaattattt
     5641 atcctgatat tataacattt tatccaaaaa aaaatatgaa tttacttttt ttatgtaaat
     5701 tgtatattta ctataaatac aaatttatat tataaaaata tattttaata atttatgaag
     5761 attaacattg taatcaatag tctttagaaa agatgggttt ctacattttt tgatttaatt
     5821 caaaatcaaa atgaaaaact atataaaatt tatttgtaat tttatatcta aaattaaaaa
     5881 aataatatgt atttttgata aaattttaat gacttaattt attttaaatt ttaagaaatt
     5941 agaattttgt aaaaattaag tgtttttgtt caaatattgc tcagaattga tttataaaat
     6001 ttaaaatatg atcattatta atatgtaagg attttcaaaa tatttatttg tacttaattt
     6061 cttattttat taattaaata tgctgatgtc taaacaaact attttttttg tttttggaaa
     6121 acttatattt gagtaattta gagaaagaca aatgcttttt ttttttgggt tttttttttt
     6181 aaatgaaatt tgttttacaa acgaatagaa agtatagatg aaggtgaaat acatatttga
     6241 atttttttta tataaaagat taaagttgtt ggttgttatg aaaggttgaa gtgaagttgg
     6301 tttgaaggtt gaagttggat gaatgaatga aaggttgaag tttggtgaat gaatgaaagg
     6361 ttgaagttgg atgaatgaaa gttgaagttt tggtttgaat gaagggtttt gtgaaagtga
     6421 tttgttcttt ctttacttga ttatttttgg ttattgttta tgagagtgga ttcgctcttg
     6481 catatgataa aaggcaaaga aagaaagata tttttaatta aggaaagccc gggggatctg
     6541 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg
     6601 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     6661 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6721 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6781 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     7021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7201 tttaacgcga attttaacaa aatattaacg cttacaattt aggtg
//
