LOCUS       Exported                6302 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 6302)
  AUTHORS   LeonardLab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 10, 2020 from SnapGene Viewer 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6302
                     /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             911..976
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             983..1003
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1028..1177
                     /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             1208..1990
                     /codon_start=1
                     /product="From Chavez et al., Nature Methods 2015"
                     /label=p65 AD
                     /translation="SQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRR
                     IAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQ
                     APAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDE
                     DLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGA
                     QRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALL"
     CDS             2009..2578
                     /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             2618..2890
                     /codon_start=1
                     /product="From Khalil et al., Cell 2012. Contains the
                     following mutations: R6A, R15A, R43A, R71A."
                     /label=ZF43(AAAA)
                     /translation="ARPGEAPFQCRICMANFSRQDRLDRHTRTHTGEKPFQCRICMANF
                     SQKEHLAGHLRTHTGEKPFQCRICMANFSRRDNLNRHLKTHLRGS"
     polyA_signal    2941..3165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3211..3639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      3709..3844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(4550..5135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5306..6166)
                     /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(6167..6271)
                     /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 ggagtcccca agctggctag
      901 cgccaccatg gattataagg atcacgatgg cgactacaag gaccacgata ttgactacaa
      961 agacgatgac gataaaatgg ctcctaagaa aaagcgcaaa gtcggtatcc atggcgttcc
     1021 cggatcagac gcattggacg attttgatct ggatatgctg ggaagtgacg ccctcgatga
     1081 ttttgacctt gacatgcttg gttcggatgc ccttgatgac tttgacctcg acatgctcgg
     1141 cagtgacgcc cttgatgatt tcgacctgga catgctgatt aactctagaa gttccggatc
     1201 tgttggtagc cagtacctgc ccgacaccga cgaccggcac cggatcgagg aaaagcggaa
     1261 gcggacctac gagacattca agagcatcat gaagaagtcc cccttcagcg gccccaccga
     1321 ccctagacct ccacctagaa gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa
     1381 acctgccccc cagccttacc ccttcaccag cagcctgagc accatcaact acgacgagtt
     1441 ccctaccatg gtgttcccca gcggccagat ctctcaggcc tctgctctgg ctccagcccc
     1501 tcctcaggtg ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg tgtctgcact
     1561 ggctcaggca ccagcacccg tgcctgtgct ggctcctgga cctccacagg ctgtggctcc
     1621 accagcccct aaacctacac aggccggcga gggcacactg tctgaagctc tgctgcagct
     1681 gcagttcgac gacgaggatc tgggagccct gctgggaaac agcaccgatc ctgccgtgtt
     1741 caccgacctg gccagcgtgg acaacagcga gttccagcag ctgctgaacc agggcatccc
     1801 tgtggcccct cacaccaccg agcccatgct gatggaatac cccgaggcca tcacccggct
     1861 cgtgacaggc gctcagaggc ctcctgatcc agctcctgcc cctctgggag caccaggcct
     1921 gcctaatgga ctgctgtctg gcgacgagga cttcagctct atcgccgata tggatttctc
     1981 agccttgctg ggctctggca gcggcagccg ggattccagg gaagggatgt ttttgccgaa
     2041 gcctgaggcc ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa
     2101 acgaatccgg ccatttcatc ctccaggaag tccatgggcc aaccgcccac tccccgccag
     2161 cctcgcacca acaccaaccg gtccagtaca tgagccagtc gggtcactga ccccggcacc
     2221 agtccctcag ccactggatc cagcgcccgc agtgactccc gaggccagtc acctgttgga
     2281 ggatcccgat gaagagacga gccaggctgt caaagccctt cgggagatgg ccgatactgt
     2341 gattccccag aaggaagagg ctgcaatctg tggccaaatg gacctttccc atccgccccc
     2401 aaggggccat ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct
     2461 ggactcaccc ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg
     2521 cctcttgcat gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgttttc
     2581 tggttctggt tctggttcag gatccggtac cggtggagct agacccgggg aggccccctt
     2641 ccagtgtcgc atttgcatgg cgaacttttc gcgccaggac aggcttgaca ggcatacccg
     2701 tactcatacc ggtgaaaaac cgtttcagtg tcggatctgt atggcaaatt tctcccagaa
     2761 ggagcacttg gcggggcatc tacgtacgca caccggcgag aagccattcc aatgccgaat
     2821 atgcatggcc aacttcagtc gccgcgacaa cctgaaccgg cacctaaaaa cccacctgag
     2881 aggatcttaa gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
     2941 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3001 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3061 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3121 gggaagtcaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3181 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3241 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3301 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3361 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3421 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3481 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3541 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3601 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3661 gttaggctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3721 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3781 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3841 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg aggttgggct
     3901 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3961 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4021 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4081 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     5101 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5161 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5221 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5281 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5341 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5401 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac
     5461 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5521 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5581 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5641 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5701 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5761 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5821 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5881 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5941 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6001 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6061 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6121 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6181 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6241 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6301 tc
//
