LOCUS       Exported                5800 bp ds-DNA     circular SYN 20-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5800)
  AUTHORS   UMC-Utrecht
  TITLE     Direct Submission
  JOURNAL   Exported Mar 8, 2018 from SnapGene Viewer 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5800
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1329
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1375..2409
                     /label=Aurora B KD +ATG
                     /note="EcoRI/EcoRI"
     misc_feature    1690..1692
                     /label=K106A
                     /note="from AAG to AGG"
     polyA_signal    2588..2709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2716..3171)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3198..3302
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3304..3661
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3696..4490
                     /product="neomycin phosphotransferase"
                     /label=KanaR/NeoR
                     /note="/vntifkey=4"
                     /db_xref="GI:1377916"
                     /protein_id="AAB02577.1"
     polyA_signal    4722..4769
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5098..5686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatctcg agctcaagct tcgaattcgc caccatggcc
     1381 cagaaggaga actcctaccc ctggccctac ggccgacaga cggctccatc tggcctgagc
     1441 accctgcccc agcgagtcct ccggaaagag cctgtcaccc catctgcact tgtcctcatg
     1501 agccgctcca atgtccagcc cacagctgcc cctggccaga aggtgatgga gaatagcagt
     1561 gggacacccg acatcttaac gcggcacttc acaattgatg actttgagat tgggcgtcct
     1621 ctgggcaaag gcaagtttgg aaacgtgtac ttggctcggg agaagaaaag ccatttcatc
     1681 gtggcgctca gggtcctctt caagtcccag atagagaagg agggcgtgga gcatcagctg
     1741 cgcagagaga tcgaaatcca ggcccacctg caccatccca acatcctgcg tctctacaac
     1801 tatttttatg accggaggag gatctacttg attctagagt atgccccccg cggggagctc
     1861 tacaaggagc tgcagaagag ctgcacattt gacgagcagc gaacagccac gatcatggag
     1921 gagttggcag atgctctaat gtactgccat gggaagaagg tgattcacag agacataaag
     1981 ccagaaaatc tgctcttagg gctcaaggga gagctgaaga ttgctgactt cggctggtct
     2041 gtgcatgcgc cctccctgag gaggaagaca atgtgtggca ccctggacta cctgccccca
     2101 gagatgattg aggggcgcat gcacaatgag aaggtggatc tgtggtgcat tggagtgctt
     2161 tgctatgagc tgctggtggg gaacccaccc tttgagagtg catcacacaa cgagacctat
     2221 cgccgcatcg tcaaggtgga cctaaagttc cccgcttccg tgcccatggg agcccaggac
     2281 ctcatctcca aactgctcag gcataacccc tcggaacggc tgcccctggc ccaggtctca
     2341 gcccaccctt gggtccgggc caactctcgg agggtgctgc ctccctctgc ccttcaatct
     2401 gtcgcctgac ccgggaatat cactagtgaa ttctgcagtc gacggtaccg cgggcccggg
     2461 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2521 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2581 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2641 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2701 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2761 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2821 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2881 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2941 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3001 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3061 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3121 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3181 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3301 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3421 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3481 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3541 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3601 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3661 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3721 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3781 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3841 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3901 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3961 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4021 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4081 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4141 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4201 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4261 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4321 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4381 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4441 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4501 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4561 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4621 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4681 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4741 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4801 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4861 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4921 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4981 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5041 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5101 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5161 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5221 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5281 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5341 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5401 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5461 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5521 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5581 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5641 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5701 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5761 atcccctgat tctgtggata accgtattac cgccatgcat
//
