LOCUS       Exported                6349 bp ds-DNA     circular SYN 08-APR-2018
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  AUTHORS   Leonard Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 10, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6349
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             910..975
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             982..1002
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1027..1176
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             1207..1989
                     /codon_start=1
                     /product="From Chavez et al., Nature Methods 2015"
                     /label=p65 AD
                     /translation="SQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRR
                     IAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQ
                     APAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDE
                     DLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGA
                     QRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALL"
     CDS             2008..2577
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             2656..2934
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="MLEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKE
                     TSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISKTR"
     polyA_signal    2988..3212
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3258..3686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3756..3891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(4597..5182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5353..6213)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6214..6318)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagtcccaa gctggctagc
      901 gccaccatgg attataagga tcacgatggc gactacaagg accacgatat tgactacaaa
      961 gacgatgacg ataaaatggc tcctaagaaa aagcgcaaag tcggtatcca tggcgttccc
     1021 ggatcagacg cattggacga ttttgatctg gatatgctgg gaagtgacgc cctcgatgat
     1081 tttgaccttg acatgcttgg ttcggatgcc cttgatgact ttgacctcga catgctcggc
     1141 agtgacgccc ttgatgattt cgacctggac atgctgatta actctagaag ttccggatct
     1201 gttggtagcc agtacctgcc cgacaccgac gaccggcacc ggatcgagga aaagcggaag
     1261 cggacctacg agacattcaa gagcatcatg aagaagtccc ccttcagcgg ccccaccgac
     1321 cctagacctc cacctagaag aatcgccgtg cccagcagat ccagcgccag cgtgccaaaa
     1381 cctgcccccc agccttaccc cttcaccagc agcctgagca ccatcaacta cgacgagttc
     1441 cctaccatgg tgttccccag cggccagatc tctcaggcct ctgctctggc tccagcccct
     1501 cctcaggtgc tgcctcaggc tcctgctcct gcaccagctc cagccatggt gtctgcactg
     1561 gctcaggcac cagcacccgt gcctgtgctg gctcctggac ctccacaggc tgtggctcca
     1621 ccagccccta aacctacaca ggccggcgag ggcacactgt ctgaagctct gctgcagctg
     1681 cagttcgacg acgaggatct gggagccctg ctgggaaaca gcaccgatcc tgccgtgttc
     1741 accgacctgg ccagcgtgga caacagcgag ttccagcagc tgctgaacca gggcatccct
     1801 gtggcccctc acaccaccga gcccatgctg atggaatacc ccgaggccat cacccggctc
     1861 gtgacaggcg ctcagaggcc tcctgatcca gctcctgccc ctctgggagc accaggcctg
     1921 cctaatggac tgctgtctgg cgacgaggac ttcagctcta tcgccgatat ggatttctca
     1981 gccttgctgg gctctggcag cggcagccgg gattccaggg aagggatgtt tttgccgaag
     2041 cctgaggccg gctccgctat tagtgacgtg tttgagggcc gcgaggtgtg ccagccaaaa
     2101 cgaatccggc catttcatcc tccaggaagt ccatgggcca accgcccact ccccgccagc
     2161 ctcgcaccaa caccaaccgg tccagtacat gagccagtcg ggtcactgac cccggcacca
     2221 gtccctcagc cactggatcc agcgcccgca gtgactcccg aggccagtca cctgttggag
     2281 gatcccgatg aagagacgag ccaggctgtc aaagcccttc gggagatggc cgatactgtg
     2341 attccccaga aggaagaggc tgcaatctgt ggccaaatgg acctttccca tccgccccca
     2401 aggggccatc tggatgagct gacaaccaca cttgagtcca tgaccgagga tctgaacctg
     2461 gactcacccc tgaccccgga attgaacgag attctggata ccttcctgaa cgacgagtgc
     2521 ctcttgcatg ccatgcatat cagcacagga ctgtccatct tcgacacatc tctgttttct
     2581 ggttctggtt ctggttcagg atccggtggt ggcgggtctg gaggaggagg tagtggtgga
     2641 ggtggctctg gtaccatgct tgagatgtgg catgaaggcc tggaagaggc atctcgtttg
     2701 tactttgggg aaaggaacgt gaaaggcatg tttgaggtgc tggagccctt gcatgctatg
     2761 atggaacggg gaccccagac tctgaaggaa acatccttta atcaggccta tggtcgagat
     2821 ttaatggagg cccaagagtg gtgcaggaag tacatgaaat cagggaatgt caaggacctc
     2881 ctccaagcct gggacctcta ttatcatgtg ttccgacgaa tctcaaagac tagatgagcg
     2941 gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3001 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3061 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3121 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagtcaatag
     3181 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3241 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3301 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3361 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3421 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3481 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3541 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3601 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3661 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt aggctcaatt
     3721 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3781 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3841 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3901 gcaaaaagct cccgggagct tgtatatcca ttttcggagg ttgggcttcg gaatcgtttt
     3961 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4021 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4081 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4141 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4201 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4261 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4321 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4381 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4441 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4501 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4561 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4621 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4681 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4741 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4801 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4861 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4921 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4981 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5041 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5101 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     5161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5281 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5341 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5401 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5461 accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt
     5521 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5581 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5641 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5701 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5761 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5821 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5881 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5941 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6001 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6061 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6121 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6181 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6241 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6301 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
