LOCUS       Exported                5443 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 5443)
  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..5443
                     /organism="synthetic DNA construct"
                     /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             1045..1719
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     CDS             1759..2031
                     /codon_start=1
                     /product="From Khalil et al., Cell 2012."
                     /label=ZF37-12
                     /translation="ARPGERPFQCRICMRNFSRNFILQRHTRTHTGEKPFQCRICMRNF
                     SDRANLRRHLRTHTGEKPFQCRICMRNFSRHDQLTRHLKTHLRGS"
     polyA_signal    2082..2306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2352..2780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2850..2985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(3691..4276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4447..5307)
                     /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(5308..5412)
                     /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 ggatcaggct ccggaagtgg ctccatggat agcactgaga acgtcatcaa gcccttcatg
     1081 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1141 ggcgagggca agccctacga gggcacccag accgccaagc tccaggtgac caagggcggc
     1201 cccctgccct tcgcctggga catcctgtcc ccccagttcc agtacggctc caaggtgtac
     1261 gtgaagcacc ccgccgacat ccccgactac aagaagctgt ccttccccga gggcttcaag
     1321 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     1381 ctccaggacg gcaccttcat ctaccacgtg aagttcatcg gcgtgaactt cccctccgac
     1441 ggccccgtaa tgcagaagaa aactctgggc tgggagccct ccaccgagcg cctgtacccc
     1501 cgcgacggcg tgctgaaggg cgagatccac aaggcgctga agctgaaggg cggcggccac
     1561 tacctggtgg agttcaagtc aatctacatg gccaagaagc ccgtgaagct gcccggctac
     1621 tactacgtgg actccaagct ggacatcacc tcccacaacg aggactacac cgtggtggag
     1681 cagtacgagc gcgccgaggc ccgccaccac ctgttccagt ctggttctgg ttctggttca
     1741 ggatccggta ccggtggagc tagacccggg gagcgcccct tccagtgtcg catttgcatg
     1801 cggaactttt cgagaaattt cattttgcaa agacataccc gtactcatac cggtgaaaaa
     1861 ccgtttcagt gtcggatctg tatgcgaaat ttctccgata gagctaattt gaggaggcat
     1921 ctgcgtacgc acaccggcga gaagccattc caatgccgaa tatgcatgcg caacttcagt
     1981 agacatgatc aattgactag acacctaaaa acccacctga gaggatctta agcggccgct
     2041 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2101 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2161 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2221 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagtca atagcaggca
     2281 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2341 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2401 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2461 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2521 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2581 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2641 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2701 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2761 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttaggctc aattagtcag
     2821 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2881 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2941 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3001 agctcccggg agcttgtata tccattttcg gaggttgggc ttcggaatcg ttttccggga
     3061 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3121 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3181 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3241 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     3301 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3361 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3421 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3481 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4201 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     4261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4561 tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc
     4621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5401 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
