LOCUS       Exported                4961 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 4961)
  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..4961
                     /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             1040..1273
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             1277..1549
                     /codon_start=1
                     /product="From Khalil et al., Cell 2012. Containst the
                     following mutations: R6A, R15A, R43A."
                     /label=ZF43(AAAR)
                     /translation="ARPGEAPFQCRICMANFSRQDRLDRHTRTHTGEKPFQCRICMANF
                     SQKEHLAGHLRTHTGEKPFQCRICMRNFSRRDNLNRHLKTHLRGS"
     polyA_signal    1600..1824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1870..2298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      2368..2503
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(3209..3794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3965..4825)
                     /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(4826..4930)
                     /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 cgtcgacggt ggactgcagg ccccaccaac cgatgtttct ttaggtgatg agttacattt
     1081 ggacggtgag gacgtagcta tggcacatgc agatgcctta gacgattttg acttggatat
     1141 gttaggagat ggagattcac caggacctgg tttcacacca cacgattcag ctccatacgg
     1201 tgctttggac atggccgatt tcgaatttga acagatgttt actgatgcct tgggtataga
     1261 cgaatacggt ggtggagcta gacccgggga ggcccccttc cagtgtcgca tttgcatggc
     1321 gaacttttcg cgccaggaca ggcttgacag gcatacccgt actcataccg gtgaaaaacc
     1381 gtttcagtgt cggatctgta tggcaaattt ctcccagaag gagcacttgg cggggcatct
     1441 acgtacgcac accggcgaga agccattcca atgccgaata tgcatgcgca acttcagtcg
     1501 ccgcgacaac ctgaaccggc acctaaaaac ccacctgaga ggatcttaag cggccgctcg
     1561 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     1621 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1681 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1741 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagtcaat agcaggcatg
     1801 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     1861 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1921 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1981 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2041 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2101 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2161 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2221 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2281 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttaggctcaa ttagtcagca
     2341 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2401 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     2461 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     2521 ctcccgggag cttgtatatc cattttcgga ggttgggctt cggaatcgtt ttccgggacg
     2581 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     2641 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2701 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2761 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     2821 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2881 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2941 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3001 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3721 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4081 gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat ttatcagcaa
     4141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
