LOCUS       Exported                7858 bp ds-DNA     circular SYN 27-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7858)
  AUTHORS   Michael Lampson
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 27, 2021 from SnapGene Viewer 5.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7858
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..3023
                     /label=RMCE_CAG-3UTR
     misc_feature    1..3023
                     /label=pEM791
     misc_feature    1..311
                     /label=TKpA
     misc_feature    312..345
                     /label=Lox2271
     CDS             350..949
                     /codon_start=1
                     /label=Pur
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    459..3023
                     /label=pEM705
     misc_feature    complement(669..688)
                     /label=EMO926 (pur_seqdn1)
     misc_feature    1091..1285
                     /label=bGH_3'UTR
     misc_feature    1097..1116
                     /label=bGH_3'UTR_seq2
     misc_feature    1181..1186
                     /label=pA
     misc_feature    1285..3000
                     /label=CAG promoter
     primer_bind     1996..2015
                     /label=EMO674
     primer_bind     complement(2086..2105)
                     /label=EMO502
     misc_feature    3002..3023
                     /label=HiLo_MCS_219-220
     misc_feature    3031..3336
                     /label=pEM791
     misc_feature    3031..3336
                     /label=pEM705
     misc_feature    3031..3336
                     /label=RMCE_CAG-3UTR
     misc_feature    3031..3336
                     /label=HiLo_MCS_219-220
     CDS             3031..3330
                     /codon_start=1
                     /label=INBox
                     /note="INCENP without its N-terminal IN box"
                     /translation="PDNYGMDLNSDDSTDDEAHPRKPIPTWARGTPLSQAIIHQYYHPP
                     NLLELFGTILPLDLEDIFKKSKPRYHKRTSSAVWNSPPLQGARVPSSLAYSLKKH"
     misc_feature    3031..3047
                     /label=634
                     /note="634"
     misc_feature    3037..3330
                     /label=IN-box
                     /note="this region is responsible for binding and 
                     activating Aurora B"
     misc_feature    3321..3336
                     /label=635
                     /note="635"
     misc_feature    3343..7858
                     /label=RMCE_CAG-3UTR
     misc_feature    3343..7858
                     /label=pEM791
     misc_feature    3343..5468
                     /label=pEM705
     misc_feature    3343..4914
                     /label=HiLo_MCS_219-220
     misc_feature    3352..4045
                     /label=mCherry
                     /note="mCherry"
     misc_feature    4046..4082
                     /label=634
                     /note="634"
     CDS             4066..4365
                     /codon_start=1
                     /label=INBox
                     /note="INCENP without its N-terminal IN box"
                     /translation="PDNYGMDLNSDDSTDDEAHPRKPIPTWARGTPLSQAIIHQYYHPP
                     NLLELFGTILPLDLEDIFKKSKPRYHKRTSSAVWNSPPLQGARVPSSLAYSLKKH"
     misc_feature    4072..4365
                     /label=IN-box
                     /note="this region is responsible for binding and 
                     activating Aurora B"
     misc_feature    4356..4379
                     /label=635
                     /note="635"
     CDS             complement(4381..4393)
                     /codon_start=1
                     /label=eDHFR
                     /note="residues 2-159"
                     /translation="F*TP"
     CDS             4393..4866
                     /codon_start=1
                     /label=eDHFR
                     /note="residues 2-159"
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             4867..4895
                     /codon_start=1
                     /label=myc
                     /note="myc"
                     /translation="EQKLISEED"
     misc_feature    4896..4899
                     /label=IRES PCR 398 399
     misc_feature    4900
                     /label=HiLo_MCS_219-220
     misc_feature    4901..4907
                     /label=HiLo_MCS_219-220
     misc_feature    complement(4902..4907)
                     /label=globin 3'UTR
     misc_feature    complement(4909..4915)
                     /label=globin 3'UTR
     misc_feature    4915..5463
                     /label=globin 3'UTR
     misc_feature    4921..5228
                     /label=insert detected by sequencing CAG_miR_3U
                     /note="insert detected by sequencing CAG_miR_3U"
     misc_feature    5486..5519
                     /label=LoxP
     misc_feature    6113..6646
                     /label=ORI
     misc_feature    complement(6872..7732)
                     /label=Bla (AmpR)
     misc_feature    complement(7733..7831)
                     /label=Bla promoter
ORIGIN
        1 aagcttaata aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
       61 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
      121 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
      181 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
      241 gtcggggcgg caggccctgc catagccact ggccccgtgg gttagggacg gggtccccca
      301 ttcgagaata aataacttcg tataaagtat cctatacgaa gttatcacca tgaccgagta
      361 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
      421 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
      481 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
      541 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
      601 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
      661 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
      721 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
      781 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
      841 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
      901 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
      961 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat ggctccgacc gaagccgacc
     1021 cgggcggccc cgccgacccc gcacccgccc ccgaggccca ccgactctag agctcgctga
     1081 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1141 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1201 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1261 ggggaggatt gggaagacaa tagcactagt tattaatagt aatcaattac ggggtcatta
     1321 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1381 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1441 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1501 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1561 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1621 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     1681 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     1741 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     1801 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     1861 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     1921 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     1981 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     2041 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg
     2101 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg
     2161 ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc
     2221 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc
     2281 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg
     2341 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc
     2401 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     2461 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     2521 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg
     2581 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
     2641 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
     2701 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
     2761 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
     2821 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     2881 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     2941 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg
     3001 ctcgaccacc ggcgcgccac cggcaccatg ccagataact acgggatgga tctgaatagc
     3061 gacgactcca ccgatgatga ggcccatccc cggaagccca tccccacctg ggcccgaggc
     3121 accccgctca gccaggctat cattcaccag tactaccacc caccgaacct tctggagctc
     3181 tttggaacca ttctcccact ggacttggag gatatcttca agaagagcaa gccccgctat
     3241 cacaagcgca ccagctctgc tgtctggaac tcaccgcccc tgcagggcgc cagggtcccc
     3301 agcagcctgg cctacagcct gaagaagcac ggatccggat ccccggtcac catggtgagc
     3361 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     3421 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     3481 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     3541 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     3601 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     3661 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     3721 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     3781 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     3841 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     3901 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     3961 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     4021 ggccgccact ccaccggcgg catggacgag ctgtataagg gtaccccaga taactacggg
     4081 atggatctga atagcgacga ctccaccgat gatgaggccc atccccggaa gcccatcccc
     4141 acctgggccc gaggcacccc gctcagccag gctatcattc accagtacta ccacccaccg
     4201 aaccttctgg agctctttgg aaccattctc ccactggact tggaggatat cttcaagaag
     4261 agcaagcccc gctatcacaa gcgcaccagc tctgctgtct ggaactcacc gcccctgcag
     4321 ggcgccaggg tccccagcag cctggcctac agcctgaaga agcacggatc caccggatcc
     4381 ctggagtcta gaatcagtct gattgcggcg ttagcggtag atcgcgttat cggcatggaa
     4441 aacgccatgc cgtggaacct gcctgccgat ctcgcctggt ttaaacgcaa caccttaaat
     4501 aaacccgtga ttatgggccg ccatacctgg gaatcaatcg gtcgtccgtt gccaggacgc
     4561 aaaaatatta tcctcagcag tcaaccgggt acggacgatc gcgtaacgtg ggtgaagtcg
     4621 gtggatgaag ccatcgcggc gtgtggtgac gtaccagaaa tcatggtgat tggcggcggt
     4681 cgcgtttatg aacagttctt gccaaaagcg caaaaactgt atctgacgca tatcgacgca
     4741 gaagtggaag gcgacaccca tttcccggat tacgagccgg atgactggga atcggtattc
     4801 agcgaattcc acgatgctga tgcgcagaac tctcacagct attgctttga gattctggag
     4861 cggcgggaac aaaaactcat ctcagaagag gatctttaat taagcgctcg aggcggccgc
     4921 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     4981 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     5041 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     5101 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     5161 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     5221 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     5281 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     5341 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     5401 tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc ctcgacctga
     5461 gcgtcgagct agcatatgga tccatataac ttcgtataat gtatgctata cgaagttatt
     5521 acgataccgt caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5581 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5641 tcaatgtatc ttaggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     5701 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5761 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5821 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5881 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5941 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6001 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6061 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6121 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6181 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6241 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6301 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6361 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6421 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6481 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6541 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6601 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6661 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6721 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6781 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6841 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6901 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6961 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7021 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7081 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7141 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7201 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7261 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7321 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7381 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7441 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7501 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7561 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7621 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7681 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7741 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7801 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgcc
//
