LOCUS       Exported                4491 bp ds-DNA     circular SYN 16-JUN-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4491)
  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..4491
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    26..34
                     /label=ZF43
                     /note="Binding site for ZF43-8 (Khalil, et al. Cell 2012)"
     misc_feature    30..38
                     /label=ZF37
                     /note="Binding site for ZF37-12 (Khalil, et al. Cell 2012)"
     misc_feature    41..49
                     /label=ZF43
                     /note="Binding site for ZF43-8 (Khalil, et al. Cell 2012)"
     misc_feature    45..53
                     /label=ZF37
                     /note="Binding site for ZF37-12 (Khalil, et al. Cell 2012)"
     misc_feature    56..64
                     /label=ZF43
                     /note="Binding site for ZF43-8 (Khalil, et al. Cell 2012)"
     misc_feature    60..68
                     /label=ZF37
                     /note="Binding site for ZF37-12 (Khalil, et al. Cell 2012)"
     misc_feature    71..79
                     /label=ZF43
                     /note="Binding site for ZF43-8 (Khalil, et al. Cell 2012)"
     misc_feature    75..83
                     /label=ZF37
                     /note="Binding site for ZF37-12 (Khalil, et al. Cell 2012)"
     misc_feature    86..94
                     /label=ZF43
                     /note="Binding site for ZF43-8 (Khalil, et al. Cell 2012)"
     misc_feature    90..98
                     /label=ZF37
                     /note="Binding site for ZF37-12 (Khalil, et al. Cell 2012)"
     misc_feature    101..109
                     /label=ZF43
                     /note="Binding site for ZF43-8 (Khalil, et al. Cell 2012)"
     misc_feature    105..113
                     /label=ZF37
                     /note="Binding site for ZF37-12 (Khalil, et al. Cell 2012)"
     promoter        123..319
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     regulatory      356..365
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
     CDS             362..1081
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1132..1356
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1402..1830
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      1900..2035
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2169..2290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2339..2355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2363..2379
                     /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(2387..2417)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2432..2453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2741..3326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3497..4357)
                     /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(4358..4462)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctg aattggagtg aggacgtgta gagtgaggac gtgcggagtg
       61 aggacgtgta gagtgaggac gtgcggagtg aggacgtgta gagtgaggac gtgccctcta
      121 gatgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
      181 aactccgccc agttccgccc attctccgcc ccatcgctga ctaatttttt ttatttatgc
      241 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
      301 aggcctaggc ttttgcaaaa agcttggcat tccggtactg ttggtaaagg ctagcgccac
      361 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      421 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      481 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      541 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
      601 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      661 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      721 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      781 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
      841 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      901 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      961 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1021 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1081 ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     1141 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1201 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1261 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagtca
     1321 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     1381 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1441 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1501 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1561 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1621 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1681 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1741 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1801 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttaggctc
     1861 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     1921 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     1981 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2041 ttttgcaaaa agctcccggg agcttgtata tccattttcg gaggttgggc ttcggaatcg
     2101 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     2161 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2221 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2281 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     2341 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     2401 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     2461 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     2521 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     2581 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2641 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     2701 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     2761 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2821 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2881 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2941 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3001 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3061 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3121 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3181 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3241 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     3301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3421 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3481 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3541 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3601 gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag
     3661 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3721 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3781 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3841 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3901 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3961 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4021 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4081 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4141 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4201 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4261 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4321 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4381 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac g
//
