LOCUS       .                       7231 bp    DNA     circular SYN 22-JUN-2021
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 22, 2021 from SnapGene Viewer 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     promoter        311..614
                     /label="CMV enhancer"
                     /note="color: #c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /ApEinfo_revcolor="#c7b0e3"
     misc_feature    820..861
                     /label="teto2x2"
                     /note="color: #f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /ApEinfo_revcolor="#f58a5e"
     misc_feature    997..1007
                     /label="Geneious type: Editing History Insertion"
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
                     /note="color: #a6acb3"
                     /ApEinfo_fwdcolor="#a6acb3"
                     /ApEinfo_revcolor="#a6acb3"
     misc_feature    997..1000
                     /label="Ligation"
                     /label="nonstandard type: ligation"
                     /note="color: #a6acb3"
                     /ApEinfo_fwdcolor="#a6acb3"
                     /ApEinfo_revcolor="#a6acb3"
     CDS             1008..1010
                     /label="Start"
                     /note="color: #992b2b"
                     /ApEinfo_fwdcolor="#992b2b"
                     /ApEinfo_revcolor="#992b2b"
                     /translation="M"
     CDS             1011..1772
                     /label="ApoE2_47-300"
                     /note="color: #0bb40b"
                     /ApEinfo_fwdcolor="#0bb40b"
                     /ApEinfo_revcolor="#0bb40b"
                     /translation="ALGRFWDYLRWVQTLSEQVQEELLSSQVTQELRALMDETMKELKA
                     YKSELEEQLTPVAEETRARLSKELQAAQARLGADMEDVCGRLVQYRGEVQAMLGQSTEE
                     LRVRLASHLRKLRKRLLRDADDLQKCLAVYQAGAREGAERGLSAIRERLGPLVEQGRVR
                     AATVGSLAGQPLQERAQAWGERLRARMEEMGSRTRDRLDEVKEQVAEVRAKLEEQAQQI
                     RLQAEAFQARLKSWFEPLVEDMQRQWAGLVEK"
     CDS             1773..1796
                     /label="Dan's SGGSGGSGG linker"
                     /note="color: #9e2273"
                     /ApEinfo_fwdcolor="#9e2273"
                     /ApEinfo_revcolor="#9e2273"
                     /translation="SGGGGSGG"
     CDS             1797..2507
                     /label="Dan's mCherry"
                     /note="color: #877fec"
                     /ApEinfo_fwdcolor="#877fec"
                     /ApEinfo_revcolor="#877fec"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKA"
     source          2514..2834
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
                     /note="color: #ffffff"
                     /ApEinfo_fwdcolor="#ffffff"
                     /ApEinfo_revcolor="#ffffff"
     source          2835..3161
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
                     /note="color: #ffffff"
                     /ApEinfo_fwdcolor="#ffffff"
                     /ApEinfo_revcolor="#ffffff"
     source          3162..3197
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
                     /note="color: #ffffff"
                     /ApEinfo_fwdcolor="#ffffff"
                     /ApEinfo_revcolor="#ffffff"
     misc_feature    2508..3203
                     /label="Geneious type: Editing History Insertion"
                     /note="Geneious type: Editing History Insertion"
                     /note="/Original_Bases="
                     /note="color: #a6acb3"
                     /ApEinfo_fwdcolor="#a6acb3"
                     /ApEinfo_revcolor="#a6acb3"
     misc_feature    2508..2513
                     /label="RTgb1a N-FKBPF36V-HA (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:j-d0hi8c2"
                     /note="color: #a6acb3"
                     /ApEinfo_fwdcolor="#a6acb3"
                     /ApEinfo_revcolor="#a6acb3"
     CDS             2514..2834
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label="F36V mutant of FK506-binding protein FKBP12 CDS"
                     /note="DmrB"
                     /note="binds synthetic ligands that do not bind wild-type
                     FKBP"
                     /note="color: #993366"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
                     /ApEinfo_fwdcolor="#993366"
                     /ApEinfo_revcolor="#993366"
     CDS             2841..3161
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label="F36V mutant of FK506-binding protein FKBP12 CDS"
                     /note="DmrB"
                     /note="binds synthetic ligands that do not bind wild-type
                     FKBP"
                     /note="color: #993366"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
                     /ApEinfo_fwdcolor="#993366"
                     /ApEinfo_revcolor="#993366"
     CDS             3168..3194
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label="HA (human influenza hemagglutinin) epitope tag CDS"
                     /note="HA"
                     /note="color: #993366"
                     /translation="YPYDVPDYA"
                     /ApEinfo_fwdcolor="#993366"
                     /ApEinfo_revcolor="#993366"
     misc_feature    3200..3203
                     /label="Ligation"
                     /label="nonstandard type: ligation"
                     /note="color: #a6acb3"
                     /ApEinfo_fwdcolor="#a6acb3"
                     /ApEinfo_revcolor="#a6acb3"
     rep_origin      3535..3841
                     /label="F1 ori"
                     /note="color: #9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /ApEinfo_revcolor="#9eafd2"
     primer_bind     complement(5073..5093)
                     /label="M13-rev"
                     /note="color: #85dae9; direction: LEFT"
                     /ApEinfo_fwdcolor="#85dae9; direction: LEFT"
                     /ApEinfo_revcolor="#85dae9; direction: LEFT"
     misc_binding    complement(5099..5121)
                     /label="LacO"
                     /note="color: #84b0dc; direction: LEFT"
                     /ApEinfo_fwdcolor="#84b0dc; direction: LEFT"
                     /ApEinfo_revcolor="#84b0dc; direction: LEFT"
     promoter        complement(5125..5155)
                     /label="lac promoter"
                     /note="color: #c7b0e3; direction: LEFT"
                     /ApEinfo_fwdcolor="#c7b0e3; direction: LEFT"
                     /ApEinfo_revcolor="#c7b0e3; direction: LEFT"
     CDS             complement(6238..6897)
                     /label="AmpR"
                     /note="color: #ffef86; direction: LEFT"
                     /ApEinfo_fwdcolor="#ffef86; direction: LEFT"
                     /ApEinfo_revcolor="#ffef86; direction: LEFT"
     source          1..7231
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /ApEinfo_fwdcolor="#ffffff"
                     /ApEinfo_revcolor="#ffffff"
     misc_feature    997..3203
                     /label="RTgb1a N-FKBPF36V-HA (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:j-d0hi8c2"
                     /note="color: #a6acb3"
                     /ApEinfo_fwdcolor="#a6acb3"
                     /ApEinfo_revcolor="#a6acb3"
     misc_feature    join(3677..7708,1..1000)
                     /label="rt1a pCDNA4-TO (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source=referenceToDocument: urn:local:.:o-d81dp1x"
                     /note="color: #a6acb3"
                     /ApEinfo_fwdcolor="#a6acb3"
                     /ApEinfo_revcolor="#a6acb3"
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 ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cgccaccatg gccctgggga
     1021 gattttggga ctatctgaga tgggtgcaga ccctgagcga gcaggtgcag gaggagctcc
     1081 tgagctctca ggtgacacag gaactgagag ccctgatgga cgagactatg aaggaactga
     1141 aggcctacaa atctgagctg gaagagcagc tgacccccgt ggctgaagag acccgcgctc
     1201 gcttgtccaa agagctccag gccgcccagg cccggctggg tgccgatatg gaggacgtct
     1261 gcggccgcct ggtccagtac cgcggggaag tgcaggctat gctggggcag tctacagagg
     1321 agctccgggt cagactggcc tctcacctgc ggaagctgag gaagaggttg ctccgcgacg
     1381 ccgacgacct ccagaagtgc ctggctgtgt accaggccgg cgcccgggaa ggcgccgagc
     1441 gcggactctc cgctatcagg gagcggctcg gccctctggt tgaacagggc agagtcaggg
     1501 ctgccactgt ggggagcctc gccggacagc ccctccagga gagagcccag gcctggggcg
     1561 agcgcctgcg agctagaatg gaggagatgg ggtccagaac ccgcgaccgc ctggacgaag
     1621 tgaaggagca ggtggccgag gtgcgtgcca aactggagga acaggcccag cagatccgcc
     1681 tccaggctga ggccttccag gccagactga agagctggtt tgagcctctt gtggaggata
     1741 tgcagcggca gtgggccggg ctggtcgaga agtccggagg cggcggatcc ggcggaatgg
     1801 tgtctaaagg cgaggaggat aacatggcta taatcaagga atttatgagg ttcaaggtgc
     1861 atatggaggg atctgtgaac ggtcacgagt tcgaaatcga gggcgaaggc gaggggcgac
     1921 cctatgaagg gacacagacg gccaaactga aggtgaccaa gggtggcccc ctgcctttcg
     1981 cctgggacat cctgtcaccc cagtttatgt acggctctaa ggcttacgtt aagcaccctg
     2041 ccgatattcc ggactacttg aaactgagct tccctgaagg gttcaagtgg gaacgggtaa
     2101 tgaattttga ggatggcggt gtagtcacag ttacccagga cagctctctt caggacggag
     2161 aatttatcta taaggttaaa cttcggggca ctaacttccc atcagacggc cccgtgatgc
     2221 agaagaaaac tatgggctgg gaagccagct cagaacgcat gtatcccgag gacggagcac
     2281 tgaagggaga aatcaagcag cgactcaagc tgaaggatgg gggacattat gatgctgagg
     2341 tcaaaaccac ctataaggcc aagaaacccg ttcagcttcc tggggcatac aacgtgaata
     2401 tcaaactgga tattacatcc cataacgaag actataccat cgtggaacag tacgagcggg
     2461 ccgagggcag acatagcaca gggggcatgg atgaattgta caaggcgact agaggcgtcc
     2521 aagtcgaaac cattagtccc ggcgatggca gaacatttcc taaaagggga caaacatgtg
     2581 tcgtccatta tacaggcatg ttggaggacg gcaaaaaggt ggacagtagt agagatcgca
     2641 ataaaccttt caaattcatg ttgggaaaac aagaagtcat taggggatgg gaggagggcg
     2701 tggctcaaat gtccgtcggc caacgcgcta agctcaccat cagccccgac tacgcatacg
     2761 gcgctaccgg acatcccgga attattcccc ctcacgctac cttggtgttt gacgtcgaac
     2821 tgttgaagct cgagactaga ggagtgcagg tggagactat ctccccagga gacgggcgca
     2881 ccttccccaa gcgcggccag acctgcgtgg tgcactacac cgggatgctt gaagatggaa
     2941 agaaagttga ttcctcccgg gacagaaaca agccctttaa gtttatgcta ggcaagcagg
     3001 aggtgatccg aggctgggaa gaaggggttg cccagatgag tgtgggtcag agagccaaac
     3061 tgactatatc tccagattat gcctatggtg ccactgggca cccaggcatc atcccaccac
     3121 atgccactct cgtcttcgat gtggagcttc taaaactgga aactagttat ccgtacgacg
     3181 taccagacta cgcataagcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     3241 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3301 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3361 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3421 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3481 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3541 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3601 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3661 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3721 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3781 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3841 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3901 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4201 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4261 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4321 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     4381 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     4441 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     4501 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     4561 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     4621 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     4681 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     4741 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     4801 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4861 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4921 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4981 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5041 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5101 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5161 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5221 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5281 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5341 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5401 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5461 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5521 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5581 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5641 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5701 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5761 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5821 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5881 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5941 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6001 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6061 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6121 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6181 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6241 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6301 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6361 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6421 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6481 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6541 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6601 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6661 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6721 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6781 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6841 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6901 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6961 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7021 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7081 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7141 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7201 cacatttccc cgaaaagtgc cacctgacgt c
//
