LOCUS       Exported                4736 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 4736)
  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..4736
                     /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             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             1052..1324
                     /codon_start=1
                     /product="From Khalil et al., Cell 2012. Contains the
                     following mutations: R6A, R15A."
                     /label=ZF43(AARR)
                     /translation="ARPGEAPFQCRICMANFSRQDRLDRHTRTHTGEKPFQCRICMRNF
                     SQKEHLAGHLRTHTGEKPFQCRICMRNFSRRDNLNRHLKTHLRGS"
     polyA_signal    1375..1599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1645..2073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2143..2278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(2984..3569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3740..4600)
                     /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(4601..4705)
                     /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 cggatcctct ggctcttcag gtaccggtgg agctagaccc ggggaggccc ccttccagtg
     1081 tcgcatttgc atggcgaact tttcgcgcca ggacaggctt gacaggcata cccgtactca
     1141 taccggtgaa aaaccgtttc agtgtcggat ctgtatgcga aatttctccc agaaggagca
     1201 cttggcgggg catctacgta cgcacaccgg cgagaagcca ttccaatgcc gaatatgcat
     1261 gcgcaacttc agtcgccgcg acaacctgaa ccggcaccta aaaacccacc tgagaggatc
     1321 ttaagcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     1381 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1441 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1501 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1561 tcaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     1621 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1681 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1741 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1801 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1861 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1921 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1981 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2041 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     2101 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2221 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2281 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggaggttg ggcttcggaa
     2341 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     2401 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2461 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2521 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     2581 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2641 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2701 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2761 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2821 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2881 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2941 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3001 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3061 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3121 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3181 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3241 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3301 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3361 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3421 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3481 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     3541 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3601 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3661 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3721 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3781 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3841 agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc
     3901 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     3961 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4021 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4081 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4141 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4201 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4261 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4321 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4381 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4441 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4501 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4561 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4621 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4681 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
