LOCUS       40924_16860        4766 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEF-F3-EGFP-F3-mcs, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4766)
  AUTHORS   Kaufman WL, Kocman I, Agrawal V, Rahn HP, Besser D, Gossen M.
  TITLE     Homogeneity and persistence of transgene expression by omitting 
            antibiotic selection in cell line isolation
  JOURNAL   Nucleic Acids Res. 36 (17), E111 (2008)
  PUBMED    18682524
REFERENCE   2  (bases 1 to 4766)
  AUTHORS   Kaufman WL, Gossen M.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-MAR-2010) Structural Biology, Helmholtz Centre for 
            Infection Research, Inhoffenstr. 7, Braunschweig 38124, Germany
REFERENCE   3  (bases 1 to 4766)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4766)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2008"; volume: "36"; issue: "17"; pages: "E111"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-MAR-2010) Structural Biology, Helmholtz Centre for Infection 
            Research, Inhoffenstr. 7, Braunschweig 38124, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4766
                     /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="FRT-3 site; mutant 3 of the flp-recombinase target
                     (FRT), the target site of yeast flp recombinase (flippase)"
     CDS             1388..2104
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    2247..2368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     2375..2422
                     /note="FRT-3 site; mutant 3 of the flp-recombinase target
                     (FRT), the target site of yeast flp recombinase (flippase)"
     primer_bind     complement(2436..2452)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2487..2505)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2531..2755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3014..3602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3776..4633)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4634..4738)
                     /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 tcctattctt caaatagtat aggaacttcg aatttccggt
     1381 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1441 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1501 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1561 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1621 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1681 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1741 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1801 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1861 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1921 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1981 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     2041 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2101 caagtccggc cggactcaga tccaccggat ctagataact gatcataatc agccatacca
     2161 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2221 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2281 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2341 gtttgtccaa actcatcaat gtatcttaac gcgtgaagtt cctattccga agttcctatt
     2401 cttcaaatag tataggaact tcggaattcc cgggggatcc actagttcta gagcggccgc
     2461 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     2521 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2581 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2641 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2701 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2761 gaggcggaaa gaaccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2821 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2881 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2941 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3001 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3061 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3121 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3181 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     3241 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3301 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3361 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3421 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     3481 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3541 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3601 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3661 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3721 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3781 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3841 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3901 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3961 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4021 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4081 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4141 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4201 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4261 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4321 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4381 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4441 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4501 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4561 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4621 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4681 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4741 catttccccg aaaagtgcca cctgac
//
