LOCUS       Exported                5411 bp ds-DNA     circular SYN 19-JUN-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    BLBP-EGFP Ali ADDgene 1-5 kb promoter version present
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5411)
  AUTHORS   VIB Trial
  TITLE     Direct Submission
  JOURNAL   Exported Freitag, 19. Jun 2015 from SnapGene 2.7.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5411
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..22
                     /note="Primer with SalI site to amplify 1,5 kb BLBP promo"
     misc_feature    7..1535
                     /note="BLBP promoter"
     misc_feature    1575..1603
                     /note="Backward BLBP primer asc site"
     CDS             1617..2336
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_site      2500..2555
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(2914..2930)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2938..2954
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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)."
     promoter        complement(2962..2992)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3007..3028
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3087..3282
                     /note="SV40 promoter"
                     /note="SV40 early promoter"
     rep_origin      3133..3268
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     polyA_signal    3288..3422
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3661..4249)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4420..5280)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5281..5385)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gtcgacgatc ctttgagaac agtgtggtag ctgctgctaa agaatcaggc aaagccagaa
       61 gttgctcaca tttaggaggc ttgtgtgctg ggggttggga ggatgactgc taaatactgg
      121 ggaagcaaaa tgagctaagt ctttacagca actgtcattt gtgagagtaa accattagaa
      181 acggacggca ggtttgtggt agattccaca aggtggtgga actagtcgca tatgtagttc
      241 agacttgaag gcacacacgt atggggacag ttctggtgtt tacgataaac aggtatgctg
      301 ccatccagct agcacagcac taggctaaag cgtactgagc ccttgtcttc cgtgggagct
      361 gcagagtggg atgcatgcgt tgtgagctga ggctcaagct gcgctggcag aagagcaggg
      421 gttgccttgt cagactccag ggtctctttc tctctgagcc tgggaaagtg ccactttatt
      481 ggatctataa agccgggggg gggggggggg gaggaatctc aaggtgaaga ggaagttcac
      541 agacccctct aacgcctcta ttagaacctt ccagctattc tctcatactt gtacactgag
      601 ctggcacaca gtataggcaa gttctattcg catcacccct ctagttcctg tctccctggt
      661 tatgcaagcc tcatatttag gtagatgtga ccttaggaaa ccaaaatatc ctttaagatc
      721 ttactaactg gttgcctgtt cagcttttcc acattgatcc tgtagccccc tcgaggaggt
      781 gaaggaaaaa aatctcctct ttgtttctct aactcattaa tgaattttaa gggcactctg
      841 taaggttcct ttcccattct ggtctggttc gtacattctg agaaacacac tgtgtttgtg
      901 ttgagagttg gctccctagc tacactgtct gtcacattga tgctctgagt agggacaggg
      961 ttcatctagg aaatatattt tcactcacac tctgtatctt ttcctagttt ggcatattct
     1021 agtctgcatt tggctctctg tttaaatata aaagaaaact aaaacacacc cttcagacgc
     1081 ctatgtctga aaaatctggc atttccgtgg gtttttcttt aaggaggcct tcatttgtaa
     1141 ccaacaccat gctctcctta aggaaatcaa tctcaatgcc ctattatcct tcccttttct
     1201 ttcctcccag tttgaggctg cagttgcctt tttttttctt atcccctgct gaacctgaaa
     1261 aaccctctct tttctacagt tttctgttcc caggccccgc tgacttcctt tagagcatgg
     1321 gggggggggg gatcaggatt gtgatgtgtg aactgggagg atcttgacct actccgctaa
     1381 cccagtggcc tgagcaaatc acaaggagga ttggagccat ctgcccagcc cctcccccac
     1441 ggcagcctgc tggaaagaga caagttagtc attcaaatga ttggcttttt gcccgcttct
     1501 tctctaaata agaaggcagc agcttctgct gaggtgcaaa gggtttttgt gaattacggt
     1561 ggtgggtaag acccgagttc ctccagttcc tgcctggcgc gccaccggtc gccaccatgg
     1621 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1681 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1741 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1801 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1861 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1921 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1981 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2041 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2101 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2161 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2221 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2281 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaggta
     2341 ccgaacatcg cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     2401 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     2461 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     2521 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     2581 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gccatatgct
     2641 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     2701 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     2761 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     2821 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
     2881 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2941 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3001 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3061 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     3121 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3181 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3241 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     3301 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3361 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3421 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3481 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3541 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3601 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3661 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3721 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3781 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3841 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3901 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3961 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4021 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4081 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4141 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5401 agtgccacct g
//
