LOCUS       40924_16855        6084 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEF-F13F14-tdTomato-dpuro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  AUTHORS   Baser B, Spehr J, Bussow K, van den Heuvel J.
  TITLE     A method for specifically targeting two independent genomic 
            integration sites for co-expression of genes in CHO cells
  JOURNAL   Methods (2015) In press
  PUBMED    26658354
REFERENCE   2  (bases 1 to 6084)
  AUTHORS   Baser B, Spehr J, Buessow K, van den Heuvel J.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2015) Structure and Function of Proteins, 
            Recombinant Protein Expression, Helmholtz Centre for Infection 
            Research, Inhoffenstrasse 7, Braunschweig, Lower Saxony 38124, 
            Germany
REFERENCE   3  (bases 1 to 6084)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6084)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods 
            (2015) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUL-2015) Structure and Function of Proteins, Recombinant 
            Protein Expression, Helmholtz Centre for Infection Research, 
            Inhoffenstrasse 7, Braunschweig, Lower Saxony 38124, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        111..1300
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     misc_recomb     1322..1369
                     /note="FRT13; synthetic variant 13 of the yeast
                     flp-recombinase target (FRT)"
     CDS             1388..2815
                     /label=tdTomato
                     /note="tandem dimeric (pseudo-monomeric) derivative of
                     DsRed (Shaner et al., 2004)"
     CDS             2816..2833
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    complement(2851..2985)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     3098..3145
                     /note="FRT14; synthetic variant 14 of the yeast
                     flp-recombinase target (FRT)"
     CDS             3165..3758
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     promoter        complement(3805..3823)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3849..4073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4332..4920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5094..5951)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5952..6056)
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cgaccgagct ttgcaaagat ggataaagtt ttaaacagag
       61 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc
      121 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      181 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      241 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      301 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      361 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacgcc
      421 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
      481 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc
      541 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      601 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      661 gtcttgtaaa tgcgggccaa gatcgatctg cacactggta tttcggtttt tggggccgcg
      721 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
      781 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
      841 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
      901 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
      961 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     1021 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     1081 ttctcgatcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1141 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1201 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1261 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattagct tgaccaagct
     1321 tgaagttcct attccgaagt tcctattctc atataagtat aggaacttcg aatttccggt
     1381 cgccaccatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg
     1441 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1501 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     1561 ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc
     1621 cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1681 gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac
     1741 gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
     1801 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
     1861 gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     1921 caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac
     1981 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc
     2041 cgagggccgc caccacctgt tcctggggca tggcaccggc agcaccggca gcggcagctc
     2101 cggcaccgcc tcctccgagg acaacaacat ggccgtcatc aaagagttca tgcgcttcaa
     2161 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2221 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     2281 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca
     2341 ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg
     2401 cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga
     2461 cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
     2521 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
     2581 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt
     2641 ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt
     2701 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2761 gcgctccgag ggccgccacc acctgttcct gtacggcatg gacgagctgt acaagcatca
     2821 tcatcaccac cactgaacgc gtatttaaat gatccagaca tgataagata cattgatgag
     2881 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2941 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3001 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3061 ctctacaaat gtggtatggc tgattagtat cacgcgtgaa gttcctattc cgaagttcct
     3121 attctatcag aagtatagga acttcggaat tcggatcttc aggcaccgag tacaagccca
     3181 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     3241 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     3301 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     3361 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     3421 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     3481 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     3541 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     3601 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     3661 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     3721 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aggatcctct agagtcgctc
     3781 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     3841 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3901 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3961 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4021 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4081 ggcggaaaga accagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4141 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4201 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4501 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     4561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4741 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4921 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4981 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5041 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5101 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5161 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5221 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5281 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5341 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5401 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5461 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5521 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5581 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5641 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5701 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5761 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5821 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5881 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5941 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6001 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6061 tttccccgaa aagtgccacc tgac
//
