LOCUS       Exported                4294 bp ds-DNA     circular SYN 26-MAR-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4294)
  TITLE     Triezenberg et al., 1988, Cousens et al., 1989
REFERENCE   2  (bases 1 to 4294)
  TITLE     Hansen et al., 2014
REFERENCE   3  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   4  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   5  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   6  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   7  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   8  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   9  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   10  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   11  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   12  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   13  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   14  (bases 1 to 4294)
  TITLE     Khalil et al., 2012, Donahue et al. 2020
REFERENCE   15  (bases 1 to 4294)
  AUTHORS   Leonard Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 28, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: ""Unpublished Work""
            SGRef: number: 2; type: ""Unpublished Work""
            SGRef: number: 3; type: ""Unpublished Work""
            SGRef: number: 4; type: ""Unpublished Work""
            SGRef: number: 5; type: ""Unpublished Work""
            SGRef: number: 6; type: ""Unpublished Work""
            SGRef: number: 7; type: ""Unpublished Work""
            SGRef: number: 8; type: ""Unpublished Work""
            SGRef: number: 9; type: ""Unpublished Work""
            SGRef: number: 10; type: ""Unpublished Work""
            SGRef: number: 11; type: ""Unpublished Work""
            SGRef: number: 12; type: ""Unpublished Work""
            SGRef: number: 13; type: ""Unpublished Work""
            SGRef: number: 14; type: ""Unpublished Work""
FEATURES             Location/Qualifiers
     source          1..4294
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    26..34
                     /label=ZF1 BS
                     /label=ZF43
                     /note="Binding site for ZF43-8"
                     /citation="[3]"
     misc_feature    30..38
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[4]"
     misc_feature    41..49
                     /label=ZF1 BS
                     /label=ZF43
                     /note="Binding site for ZF43-8"
                     /citation="[5]"
     misc_feature    45..53
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[6]"
     misc_feature    56..64
                     /label=ZF1 BS
                     /label=ZF43
                     /note="Binding site for ZF43-8"
                     /citation="[7]"
     misc_feature    60..68
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[8]"
     misc_feature    71..79
                     /label=ZF1 BS
                     /label=ZF43
                     /note="Binding site for ZF43-8"
                     /citation="[9]"
     misc_feature    75..83
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[10]"
     misc_feature    86..94
                     /label=ZF1 BS
                     /label=ZF43
                     /note="Binding site for ZF43-8"
                     /citation="[11]"
     misc_feature    90..98
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[12]"
     misc_feature    101..109
                     /label=ZF1 BS
                     /label=ZF43
                     /note="Binding site for ZF43-8"
                     /citation="[13]"
     misc_feature    105..113
                     /label=ZF2 BS
                     /note="Binding site for ZF37-12"
                     /citation="[14]"
     promoter        117..141
                     /label=YB_TATA Minimal Promoter
                     /citation="[2]"
     CDS             177..245
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     CDS             252..272
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             309..542
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes
                     simplex virus protein VP16"
                     /label=VP16 AD
                     /citation="[1]"
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             543..602
                     /codon_start=1
                     /label=Linker
                     /translation="GGSGGGGSGGGGSGGGGSGT"
     CDS             603..881
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="MLEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKE
                     TSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISKTR"
     polyA_signal    935..1159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1205..1633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      1703..1838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1972..2093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2166..2182
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(2190..2220)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2235..2256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2544..3129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3303..4160)
                     /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(4161..4265)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctg aattggagtg aggacgtgta gagtgaggac gtgcggagtg
       61 aggacgtgta gagtgaggac gtgcggagtg aggacgtgta gagtgaggac gtgccctcta
      121 gagggtatat aatgggggcc actagtctac taccagagct catcgctagc gccaccatgg
      181 attataagga tcacgatggc gactacaagg accacgatat tgactacaaa gacgatgacg
      241 ataaaatggc tcctaagaaa aagcgcaaag tcggtatcca tggcgttccc gtcgacggtg
      301 gactccaagc cccaccaacc gatgtttctt taggtgatga gttacatttg gacggtgagg
      361 acgtagctat ggcacatgca gatgccttag acgattttga cttggatatg ttaggagatg
      421 gagattcacc aggacctggt ttcacaccac acgattcagc tccatacggt gctttggaca
      481 tggccgattt cgaatttgaa cagatgttta ctgatgcctt gggtatagac gaatacggtg
      541 gtggaggatc cggtggtggc gggtctggag gaggaggtag tggtggaggt ggctctggta
      601 ccatgcttga gatgtggcat gaaggcctgg aagaggcatc tcgtttgtac tttggggaaa
      661 ggaacgtgaa aggcatgttt gaggtgctgg agcccttgca tgctatgatg gaacggggac
      721 cccagactct gaaggaaaca tcctttaatc aggcctatgg tcgagattta atggaggccc
      781 aagagtggtg caggaagtac atgaaatcag ggaatgtcaa ggacctcctc caagcctggg
      841 acctctatta tcatgtgttc cgacgaatct caaagactag atgagcggcc gctcgagtct
      901 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
      961 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1021 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1081 ggtggggtgg ggcaggacag caagggggag gattgggaag tcaatagcag gcatgctggg
     1141 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     1201 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1261 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1321 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1381 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1441 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1501 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1561 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1621 tttaacgcga attaattctg tggaatgtgt gtcagttagg ctcaattagt cagcaaccat
     1681 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1741 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     1801 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     1861 gggagcttgt atatccattt tcggaggttg ggcttcggaa tcgttttccg ggacgccggc
     1921 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     1981 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2041 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2101 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2161 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2221 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2281 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2341 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2401 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2461 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2521 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2581 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2641 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2701 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2761 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2821 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2881 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2941 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3001 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3061 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3121 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3181 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3241 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3301 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3361 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3421 agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac
     3481 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3541 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3601 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3661 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3721 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3781 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3841 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3901 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3961 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4021 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4081 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4141 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4201 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4261 ccgcgcacat ttccccgaaa agtgccacct gacg
//
