LOCUS       40924_17209        6125 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Transformation vector pEGFP-neo4 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6125)
  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 6125)
  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 6125)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6125)
  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..6125
                     /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..2963
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2970..3368
                     /label=BTU1 3' flanking
                     /note="BTU1 3' flanking"
     CDS             4280..5071
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5440..5456)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5482..5500)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5507..5523)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5665..6120
                     /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 tttctaaggg tgaagaactt ttcactggtg
     2281 ttgttcctat tttagttgaa ttagatggtg atgttaatgg tcacaaattt tctgtttcag
     2341 gtgaaggtga aggtgatgct acttatggta agttaactct taaatttatt tgtactactg
     2401 gtaagttacc tgttccttgg cctactttag ttactactct tacttatggt gttcaatgct
     2461 tttcaagata ccctgatcac atgaagcaac atgatttctt taagtctgct atgcctgaag
     2521 gttatgttca agaaagaact attttcttta aagatgatgg taattataaa actagagctg
     2581 aagttaaatt tgaaggtgat actttagtta atagaattga acttaaaggt attgatttca
     2641 aggaagatgg taatattctt ggtcataaac ttgaatataa ttacaattct cataatgttt
     2701 acattatggc tgataagcaa aagaatggta ttaaagttaa ttttaaaatt agacataata
     2761 ttgaagatgg ttcagttcaa ttagctgatc attatcaaca aaatactcct attggtgatg
     2821 gtcctgttct tttacctgat aatcactact tatctactca atcagcttta tctaaagatc
     2881 ctaatgaaaa aagagatcat atggttcttt tagaatttgt tactgctgct ggtattactc
     2941 ttggtatgga tgaattatat aagactagtt gagcgaactg aatcggtcag ctaaaccaac
     3001 caatcaacat aataaacttt attattttta cttaagcatc ttactgttgt tgtaatagta
     3061 gagaaagaaa tacccaatta acttcattca cataacatta atatctataa acatcttttt
     3121 tctcacatat atacaactct ctaaatcaac aaataacttt ttaaaaataa tggatatata
     3181 ttaacaaaat aatatatctc tttttacaaa atagttctta tataaatacg tattctgcac
     3241 tcacccgcat ttttcacaac aaaaacatac caaaaaaatt cttacttcta catgtttcct
     3301 ttcttattat tacaaaatta ttttataaat agcatacaaa aataaataca ataaaaaaat
     3361 aaacaaaatc cagatccccc gggctgcagg aattcgatag acaatttatt tctaaaaaat
     3421 atttaaaaat aaaaaataat aagggttttg aataactcct ttaatttaaa tacacatttt
     3481 taaatttttt ttagctcttt aaatattcat aaaaataaaa aataactaac taaaaataaa
     3541 taaaaagata ataatgatta aaggtataat actgaataag aaaaaacata atagagtact
     3601 tattttttat atcactattt ttaatatctt gaaagcaaaa cttttttata tatcttaaaa
     3661 tatattgtat cgtttattca attattttct ttaaatttca aatatattga taaaaaagat
     3721 gacatgtttt ttaaagaaaa catgaaatat aaaatagata aatatcaatt attttattta
     3781 ttaaatatat aagctgctca aaacatagct cattcatcaa ttataatatg tgaatcatta
     3841 attttcaaaa tattactcat tatttaggct atcatttatt ttttattttc aattatccgt
     3901 ttctattata ttttaatatt aagttgtgat tcttgaattt tgtgtcatga attatttgta
     3961 aatcttttta tttctgataa aaaatataaa ttgattgact catgatttaa atcatgagtc
     4021 aacctaacta attttcaaaa ttcttctatt ctaaaatata gatgtgattc ttgaatctct
     4081 cttgaatata aagtaatttt ttatatttct gatataattc ttagctacgt gattcacgat
     4141 ttatacaatg atccatataa aataatgtaa atagtgtata tatatatatt cgtctttttt
     4201 attctttata taatttaaaa aaattaaaaa aatttaataa agctctaata aaataaataa
     4261 taatactaaa cttaaaataa tgattgaaca agatggttta cacgctggtt ctcccgccgc
     4321 ttgggtcgaa agacttttcg gttatgactg ggctcaacaa accatcggtt gctctgatgc
     4381 cgccgtcttc cgtctttctg ctcaaggtcg tcctgttctt ttcgtcaaga ccgacctttc
     4441 tggtgccctt aatgaacttc aagatgaagc tgcccgtctt tcttggcttg ccaccaccgg
     4501 tgttccttgc gctgctgtcc ttgacgttgt cactgaagcc ggtagagact ggcttctttt
     4561 aggtgaagtc cccggtcaag atcttctttc ttctcacctt gctcctgccg aaaaagtttc
     4621 tatcatggct gatgctatgc gtcgtcttca tacccttgat cccgctacct gccctttcga
     4681 ccaccaagcc aaacatcgta tcgaacgtgc tcgtactcgt atggaagccg gtcttgtcga
     4741 tcaagatgat cttgacgaag aacatcaagg tcttgcccct gccgaacttt tcgccagact
     4801 taaggcccgt atgcccgacg gtgaagatct tgtcgtcacc catggtgatg cctgcttacc
     4861 caatatcatg gttgaaaatg gtcgtttttc tggtttcatc gactgtggtc gtcttggtgt
     4921 cgccgaccgt tatcaagata ttgccttagc tacccgtgat attgctgaag aacttggtgg
     4981 tgaatgggct gaccgtttcc ttgtccttta cggtatcgcc gctcccgatt ctcaacgtat
     5041 cgccttctat cgtcttcttg acgaattctt ctgagatcct taaattaaaa attcaatata
     5101 tatttacaaa ctttcatata aaataaatat attatataaa attaattttt agtgtattat
     5161 attaacatta aagcaccaaa aaaacgtgtt aatatactac tataaaatat aatttattcc
     5221 aaattgacta aaatcattat tttacaactc atttgtatat atattttatg tcaattattt
     5281 tttttaactt tctaaaaaaa aaaattcctc ttcacataca tgttagctct taaaaatttg
     5341 tctgcaaatc caataataat attttttttt gccattaaat tttcaaattt ttactggaaa
     5401 aatgcagccc gggggatctg aattcgatat caagcttatc gataccgtcg acctcgaggg
     5461 ggggcccggt acccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt
     5521 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5581 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5641 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5701 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5761 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5821 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5881 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5941 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6001 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6061 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6121 aggtg
//
