LOCUS       40924_30001        6116 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Transformation vector pmCherry-neo4 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6116)
  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 6116)
  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 6116)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6116)
  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..6116
                     /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..2954
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    2961..3359
                     /label=BTU1 3' flanking
                     /note="BTU1 3' flanking"
     CDS             4271..5062
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5431..5447)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5473..5491)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5498..5514)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5656..6111
                     /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 ggatccatgg tttcaaaagg agaagaagat aacatggcta
     2281 ttattaaaga atttatgaga tttaaggttc atatggaagg ttctgttaac ggtcacgaat
     2341 ttgaaattga aggagaagga gaaggtagac cttatgaagg aacacaaact gctaaattaa
     2401 aagtaactaa aggtggtcct ttacctttcg catgggatat tctttcacct caatttatgt
     2461 atggttctaa agcatatgtt aaacaccctg ctgatattcc tgattattta aagttgtctt
     2521 ttcctgaagg ttttaagtgg gaaagagtta tgaattttga agatggtgga gttgttacag
     2581 taactcaaga ttcttcttta caagatggag aatttattta taaagttaaa ttaagaggta
     2641 ctaattttcc tagcgatgga cctgtcatgc aaaagaaaac aatgggttgg gaagcaagtt
     2701 ctgaaagaat gtaccctgaa gatggtgcac ttaaaggaga aattaagcaa agattaaaac
     2761 ttaaagatgg tggacattat gatgctgaag tcaaaactac ttataaagca aagaaacctg
     2821 ttcaattacc tggtgcatac aatgttaata ttaagttaga tattacatct cataatgaag
     2881 attacacaat agtagaacaa tacgaaagag ctgaaggtag acacagtact ggtggtatgg
     2941 atgaacttta caaaactagt tgagcgaact gaatcggtca gctaaaccaa ccaatcaaca
     3001 taataaactt tattattttt acttaagcat cttactgttg ttgtaatagt agagaaagaa
     3061 atacccaatt aacttcattc acataacatt aatatctata aacatctttt ttctcacata
     3121 tatacaactc tctaaatcaa caaataactt tttaaaaata atggatatat attaacaaaa
     3181 taatatatct ctttttacaa aatagttctt atataaatac gtattctgca ctcacccgca
     3241 tttttcacaa caaaaacata ccaaaaaaat tcttacttct acatgtttcc tttcttatta
     3301 ttacaaaatt attttataaa tagcatacaa aaataaatac aataaaaaaa taaacaaaat
     3361 ccagatcccc cgggctgcag gaattcgata gacaatttat ttctaaaaaa tatttaaaaa
     3421 taaaaaataa taagggtttt gaataactcc tttaatttaa atacacattt ttaaattttt
     3481 tttagctctt taaatattca taaaaataaa aaataactaa ctaaaaataa ataaaaagat
     3541 aataatgatt aaaggtataa tactgaataa gaaaaaacat aatagagtac ttatttttta
     3601 tatcactatt tttaatatct tgaaagcaaa acttttttat atatcttaaa atatattgta
     3661 tcgtttattc aattattttc tttaaatttc aaatatattg ataaaaaaga tgacatgttt
     3721 tttaaagaaa acatgaaata taaaatagat aaatatcaat tattttattt attaaatata
     3781 taagctgctc aaaacatagc tcattcatca attataatat gtgaatcatt aattttcaaa
     3841 atattactca ttatttaggc tatcatttat tttttatttt caattatccg tttctattat
     3901 attttaatat taagttgtga ttcttgaatt ttgtgtcatg aattatttgt aaatcttttt
     3961 atttctgata aaaaatataa attgattgac tcatgattta aatcatgagt caacctaact
     4021 aattttcaaa attcttctat tctaaaatat agatgtgatt cttgaatctc tcttgaatat
     4081 aaagtaattt tttatatttc tgatataatt cttagctacg tgattcacga tttatacaat
     4141 gatccatata aaataatgta aatagtgtat atatatatat tcgtcttttt tattctttat
     4201 ataatttaaa aaaattaaaa aaatttaata aagctctaat aaaataaata ataatactaa
     4261 acttaaaata atgattgaac aagatggttt acacgctggt tctcccgccg cttgggtcga
     4321 aagacttttc ggttatgact gggctcaaca aaccatcggt tgctctgatg ccgccgtctt
     4381 ccgtctttct gctcaaggtc gtcctgttct tttcgtcaag accgaccttt ctggtgccct
     4441 taatgaactt caagatgaag ctgcccgtct ttcttggctt gccaccaccg gtgttccttg
     4501 cgctgctgtc cttgacgttg tcactgaagc cggtagagac tggcttcttt taggtgaagt
     4561 ccccggtcaa gatcttcttt cttctcacct tgctcctgcc gaaaaagttt ctatcatggc
     4621 tgatgctatg cgtcgtcttc atacccttga tcccgctacc tgccctttcg accaccaagc
     4681 caaacatcgt atcgaacgtg ctcgtactcg tatggaagcc ggtcttgtcg atcaagatga
     4741 tcttgacgaa gaacatcaag gtcttgcccc tgccgaactt ttcgccagac ttaaggcccg
     4801 tatgcccgac ggtgaagatc ttgtcgtcac ccatggtgat gcctgcttac ccaatatcat
     4861 ggttgaaaat ggtcgttttt ctggtttcat cgactgtggt cgtcttggtg tcgccgaccg
     4921 ttatcaagat attgccttag ctacccgtga tattgctgaa gaacttggtg gtgaatgggc
     4981 tgaccgtttc cttgtccttt acggtatcgc cgctcccgat tctcaacgta tcgccttcta
     5041 tcgtcttctt gacgaattct tctgagatcc ttaaattaaa aattcaatat atatttacaa
     5101 actttcatat aaaataaata tattatataa aattaatttt tagtgtatta tattaacatt
     5161 aaagcaccaa aaaaacgtgt taatatacta ctataaaata taatttattc caaattgact
     5221 aaaatcatta ttttacaact catttgtata tatattttat gtcaattatt ttttttaact
     5281 ttctaaaaaa aaaaattcct cttcacatac atgttagctc ttaaaaattt gtctgcaaat
     5341 ccaataataa tatttttttt tgccattaaa ttttcaaatt tttactggaa aaatgcagcc
     5401 cgggggatct gaattcgata tcaagcttat cgataccgtc gacctcgagg gggggcccgg
     5461 tacccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     5521 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5581 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5641 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5701 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5761 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5821 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5881 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5941 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6001 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6061 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtg
//
