LOCUS       Exported                6435 bp ds-DNA     circular SYN 02-DEC-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pDRR-donor-BFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6435)
  AUTHORS   Nausica
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 12, 2017 from SnapGene 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6435
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             31..828
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             829..882
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /note="T2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             889..1596
                     /codon_start=1
                     /note="mCherry"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     CDS             1598..2312
                     /codon_start=1
                     /note="eGFP(opt)"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     promoter        2440..2651
                     /note="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2714..3415
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /note="TagBFP"
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_site      3523..3578
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(3939..3955)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3963..3979
                     /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(3987..4017)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4032..4053
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4112..4307
                     /note="SV40 promoter"
                     /note="SV40 early promoter"
     rep_origin      4158..4293
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     polyA_signal    4313..4447
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4686..5274)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5445..6305)
                     /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(6306..6410)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 ggtcgacatt gattagtcga ctctcagtac ggatcggcca ttgaacaaga tggattgcac
       61 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
      121 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
      181 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
      241 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
      301 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
      361 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
      421 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
      481 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
      541 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
      601 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
      661 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
      721 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
      781 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttcga gggcagagga
      841 agtcttctaa catgcggtga cgtggaggag aatccgggcc ccattagggt ctctaaagga
      901 gaggaggata atatggcaat tatcaaggag tttatgagat ttaaggtgca catggaaggc
      961 tccgtgaacg gccacgagtt cgagattgag ggagagggcg agggacggcc ctatgaggga
     1021 acacagaccg ccaagctgaa ggtcaccaag ggaggacccc tgcctttcgc atgggacatc
     1081 ctttcacctc agttcatgta cgggagtaaa gcgtatgtaa aacatccagc cgatattccc
     1141 gactatctta aactgtcctt tcccgaaggt tttaagtggg agagagtgat gaattttgaa
     1201 gatgggggcg ttgtgacggt tacacaggat tcttcactcc aagatggcga gtttatatac
     1261 aaggttaagc tgcgaggcac aaattttcct tccgatggcc cagtcatgca gaaaaaaact
     1321 atggggtggg aggcaagctc agagcgaatg taccccgaag atggggccct gaaaggagaa
     1381 attaagcaaa gactgaaact gaaggatggc ggacactacg acgccgaggt caagacaacg
     1441 tataaggcca aaaaacctgt gcagctcccg ggcgcctata acgtgaatat caagcttgac
     1501 ataacctctc acaacgaaga ctacactatc gtggagcagt acgagcgcgc tgagggaagg
     1561 catagcaccg gagggatgga cgagttgtat aagtgagggt gagcaagggc gaagaactct
     1621 ttacaggcgt cgttcccatc ttggttgagc ttgacgggga cgtcaacggc cataaattca
     1681 gcgtttccgg ggagggagag ggcgatgcaa catacgggaa gctgactttg aagtttatct
     1741 gtacgactgg aaaactgccg gtgccttggc cgacgctggt gactactctg acatatggtg
     1801 tgcagtgttt ttccagatat cctgatcata tgaagcagca tgatttcttc aagagcgcga
     1861 tgccagaggg ctacgtgcag gaaagaacga tcttcttcaa agacgatggt aattacaaaa
     1921 ctcgggcaga ggtcaaattc gagggagata cgctggtaaa ccggatcgag ctgaagggca
     1981 tcgattttaa ggaagatggt aacatcctgg gacacaaact ggagtataat tataactccc
     2041 ataatgtcta tattatggct gacaagcaga agaacgggat caaggtgaac tttaagatcc
     2101 ggcacaatat cgaagatggc tccgttcagc tggccgatca ttatcaacaa aacacaccta
     2161 ttggagatgg gccagtcctg ttgccagata atcactatct gtccacacag tccgccttgt
     2221 ccaaagaccc gaacgagaag agagaccaca tggtgctgct ggagtttgtt acggcggccg
     2281 ggatcacgtt gggcatggat gagctttaca agtgaagcgg ccgcgactca taatcagcca
     2341 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2401 gaaacataaa atgaatgcaa ttgaatcaac ctctggattg ggcagagcgc acatcgccca
     2461 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc
     2521 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     2581 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     2641 ccagaacaca gctctggcta actagagaac ccactgtagt gaaccgtcag atcgcctgga
     2701 gacgccatcc acgatgtccg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     2761 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     2821 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     2881 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     2941 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     3001 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     3061 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     3121 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     3181 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     3241 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     3301 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     3361 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta attgaatctc
     3421 atgctggagt tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     3481 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     3541 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     3601 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     3661 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     3721 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     3781 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     3841 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgaa
     3901 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     3961 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4021 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4081 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     4141 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4201 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4261 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     4321 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4381 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4441 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4501 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4561 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4621 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4861 ggaagcgtgg cgctttctca aagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5101 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5281 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5761 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6061 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6421 cgaaaagtgc cacct
//
