LOCUS       Exported                9736 bp ds-DNA     circular SYN 14-SEP-2022
DEFINITION  5 fragments assembled using overlaps.
ACCESSION   .
VERSION     .
KEYWORDS    Assembly CD shuffled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 14, 2022 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9736
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..502
                     /label=CMV'
     CDS             575..4495
                     /codon_start=1
                     /label=Ki67 shuffled Nterm
                     /label=CD_shuffled-LR-mNeonGreen
                     /translation="MASFGGHLRPELFDENLPPNTPLKRGEAPTKRKSLVMHTPPVLKK
                     IIKEQPQPSGKQESGSEIHVEVKAQSLVISPPAPSPRKTPVASDQRRRSCKTAPASSSK
                     SQTEVPKRGGRKSGNLPSKRVSISRSQHDILQMICSKRRSGASEANLIVAKSWADVVKL
                     GAKQTQTKVIKHGPQRSMNKRQRRPATPKKPVGEVHSQFSTGHANSPCTIIIGKAHTEK
                     EIHEQEAILHNFSSTNPTQVNGSVIDEPVRLKHGDVITIIDRSFRYENESLQNGRKSTE
                     FPRKIREQEPARRVSRSSFSSDPDEKAQDSKAYSKITEGKVSGNPQVHIKNVKEDSTAD
                     DSKDSVAQGTTNVHSSEHAGRNGRNAADPISGDFKEISSVKLVSRYGELKSVPTTQCLD
                     NSKKNESPFWKLYESVKKELDVKSQKENVLQYCRKSGLQTDYATEKESADGLQGETQLL
                     VSRKSRPKSGGSGHAVAEPASPEQELDQNKGKGRDVESVQTPSKAVGASFPLYEPAKMK
                     TPVQYSQQQNSPQKHKNKDLYTTGRRESVNLGKSEGFKAGDKTLTPRKLSTRNRTPAKV
                     EDAADSATKPENLSSKTRGSIPTDVEVLPTETEIHNEPFLTLWLTQVERKIQKDSLSKP
                     EKLGTTAGQMCSVLPGLSSVDINNFGDSINESEGIPLKRRRVVHVPARPYRVLNNFISN
                     QKMDFKEDLSGIAEMFKTPVKEQPQLTSTCHIAISNSENLLGKQFQGTDSGEEPLLPTS
                     ESFGGNVFFSAQNAAKQPSDKCSASPPLRRQCIRENGNVAKTPRNTYKMTSLETKTSDT
                     ETEPSKTVSTANRSGRSTEFRNIQKLPVESKSEETNTEIVECILKRGQKATLLQQRREG
                     EMKEIERPFETYKENIELKENDEKMKAMKRSRTWGQKCAPMSDLTDLKSLPDTELMKDT
                     ARGQNLLQTQDHAKAWPTRRLVTIKRSGVDGPHFPLSLSTCLFGRGIECDIRIQLPVVS
                     KQHCKMCAPKQTPDSGKPLKISRRVLRAPKVEPVGDVVSTRDPVKSQSKSNTSLPPLPF
                     KRGGGKDGSVTGTKRLRCMPAPEEIVEELPASKKQRVAPRARGKSSEPVVIMKRSLRTS
                     AKRIEPAEELNSNDMKTNKEEHKLQDSVPENKGISLRSRRQNKTEAEQQITEVFVLAER
                     IEINRNEKKPMKTSPEMDIQNPDDGARKPIPRDKVTENKRCLRSARQNESSQPKVAEES
                     GGQKSAKVLMQNQKGKGEAGNSDSMCLRSRKTKSQPAASTLESKSVQRVTRSVKRCAEN
                     PKKAEDNVCVKKIRTRSHRDSED"
     CDS             3560..4495
                     /codon_start=1
                     /gene="'Ki67_NoRepeat"
                     /label='Ki67_NoRepeat
                     /translation="CAPKQTPDSGKPLKISRRVLRAPKVEPVGDVVSTRDPVKSQSKSN
                     TSLPPLPFKRGGGKDGSVTGTKRLRCMPAPEEIVEELPASKKQRVAPRARGKSSEPVVI
                     MKRSLRTSAKRIEPAEELNSNDMKTNKEEHKLQDSVPENKGISLRSRRQNKTEAEQQIT
                     EVFVLAERIEINRNEKKPMKTSPEMDIQNPDDGARKPIPRDKVTENKRCLRSARQNESS
                     QPKVAEESGGQKSAKVLMQNQKGKGEAGNSDSMCLRSRKTKSQPAASTLESKSVQRVTR
                     SVKRCAENPKKAEDNVCVKKIRTRSHRDSED"
     misc_feature    3764
                     /label=silent mutation 2
                     /note="C -> T mutation"
     CDS             4508..5218
                     /codon_start=1
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     misc_feature    5251..5546
                     /label=IVS
     misc_feature    5572..6182
                     /label=IRES
     misc_feature    6183..6776
                     /label=Puro
     misc_feature    8432..9363
                     /label=Amp'
ORIGIN
        1 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
       61 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      121 caatgggtgg actatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      181 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      241 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      301 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      361 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      421 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      481 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      541 taccggactc agatctcgag ctcaagcttc caccatggct tcctttggtg ggcacctaag
      601 acctgaacta tttgatgaaa acttgcctcc taatacgcct ctcaaaaggg gagaagcccc
      661 aaccaaaaga aagtctctgg taatgcacac tccacctgtc ctgaagaaaa tcatcaagga
      721 acagcctcaa ccatcaggaa aacaagagtc aggttcagaa atccatgtgg aagtgaaggc
      781 acaaagcttg gttataagcc ctccagctcc tagtcctagg aaaactccag ttgccagtga
      841 tcaacgccgt aggtcctgca aaacagcccc tgcttccagc agcaaatctc agacagaggt
      901 tcctaagaga ggagggagaa agagtggcaa cctgccttca aagagagtgt ctatcagccg
      961 aagtcaacat gatattttac agatgatatg ttccaaaaga agaagtggtg cttcggaagc
     1021 aaatctgatt gttgcaaaat catgggcaga tgtagtaaaa cttggtgcaa aacaaacaca
     1081 aactaaagtc ataaaacatg gtcctcaaag gtcaatgaac aaaaggcaaa gaagacctgc
     1141 tactccaaag aagcctgtgg gcgaagttca cagtcaattt agtacaggcc acgcaaactc
     1201 tccttgtacc ataataatag ggaaagctca tactgaaaaa gaaatccatg agcaggaggc
     1261 aatattacat aatttcagtt ccacaaatcc aacacaagta aatgggtctg ttattgatga
     1321 gcctgtacgg ctaaaacatg gagatgtaat aactattatt gatcgttcct tcaggtatga
     1381 aaatgaaagt cttcagaatg gaaggaagtc aactgaattt ccaagaaaaa tacgtgaaca
     1441 ggagccagca cgtcgtgtct caagatctag cttctcttct gaccctgatg agaaagctca
     1501 agattccaag gcctattcaa aaatcactga aggaaaagtt tcaggaaatc ctcaggtaca
     1561 tatcaagaat gtcaaagaag acagtaccgc agatgactca aaagacagtg ttgctcaggg
     1621 aacaactaat gttcattcct cagaacatgc tggacgtaat ggcagaaatg cagctgatcc
     1681 catttctggg gattttaaag aaatttccag cgttaaatta gtgagccgtt atggagaatt
     1741 gaagtctgtt cccactacac aatgtcttga caatagcaaa aaaaatgaat ctcccttttg
     1801 gaagctttat gagtcagtga agaaagagtt ggatgtaaaa tcacaaaaag aaaatgtcct
     1861 acagtattgt agaaaatctg gattacaaac tgattacgca acagagaaag aaagtgctga
     1921 tggtttacag ggggagaccc aactgttggt ctcgcgtaag tcaagaccaa aatctggtgg
     1981 gagcggccac gctgtggcag agcctgcttc acctgaacaa gagcttgacc agaacaaggg
     2041 gaagggaaga gacgtggagt ctgttcagac tcccagcaag gctgtgggcg ccagctttcc
     2101 tctctatgag ccggctaaaa tgaagacccc tgtacaatat tcacagcaac aaaattctcc
     2161 acaaaaacat aagaacaaag acctgtatac tactggtaga agagaatctg tgaatctggg
     2221 taaaagtgaa ggcttcaagg ctggtgataa aactcttact cccaggaagc tttcaactag
     2281 aaatcgaaca ccagctaaag ttgaagatgc agctgactct gccactaagc cagaaaatct
     2341 ctcttccaaa accagaggaa gtattcctac agatgtggaa gttctgccta cggaaactga
     2401 aattcacaat gagccatttt taactctgtg gctcactcaa gttgagagga agatccaaaa
     2461 ggattccctc agcaagcctg agaaattggg cactacagct ggacagatgt gctctgtgtt
     2521 acctggtctt agttcagttg atatcaacaa ctttggtgat tccattaatg agagtgaggg
     2581 aatacctttg aaaagaaggc gtgtggtaca tgtgcctgct cgaccctaca gagtgctcaa
     2641 caacttcatt tccaaccaaa aaatggactt taaggaagat ctttcaggaa tagctgaaat
     2701 gttcaagacc ccagtgaagg agcaaccgca gttgacaagc acatgtcaca tcgctatttc
     2761 aaattcagag aatttgcttg gaaaacagtt tcaaggaact gattcaggag aagaacctct
     2821 gctccccacc tcagagagtt ttggaggaaa tgtgttcttc agtgcacaga atgcagcaaa
     2881 acagccatct gataaatgct ctgcaagccc tcccttaaga cggcagtgta ttagagaaaa
     2941 tggaaacgta gcaaaaacgc ccaggaacac ctacaaaatg acttctctgg agacaaaaac
     3001 ttcagatact gagacagagc cttcaaaaac agtatccact gcaaacaggt caggaaggtc
     3061 tacagagttc aggaatatac agaagctacc tgtggaaagt aagagtgaag aaacaaatac
     3121 agaaattgtt gagtgcatcc taaaaagagg tcagaaggca acactactac aacaaaggag
     3181 agaaggagag atgaaggaaa tagaaagacc ttttgagaca tataaggaaa atattgaatt
     3241 aaaagaaaac gatgaaaaga tgaaagcaat gaagagatca agaacttggg ggcagaaatg
     3301 tgcaccaatg tctgacctga cagacctcaa gagcttgcct gatacagaac tcatgaaaga
     3361 cacggcacgt ggccagaatc tcctccaaac ccaagatcat gccaaggcat ggcccacgag
     3421 acgcctggtt actatcaaaa ggagcggggt cgacggtccc cactttcccc tgagcctcag
     3481 cacctgcttg tttggaaggg gtattgaatg tgacatccgt atccagcttc ctgttgtgtc
     3541 aaaacaacat tgcaaaatgt gcgctccaaa gcaaacacct gacagtggaa aacctctaaa
     3601 aatatccaga agagttcttc gggcccctaa agtagaaccc gtgggagacg tggtaagcac
     3661 cagagaccct gtaaaatcac aaagcaaaag caacacttcc ctgcccccac tgcccttcaa
     3721 gaggggaggt ggcaaagatg gaagcgtcac gggaaccaag aggttgcgct gcatgccagc
     3781 accagaggaa attgtggagg agctgccagc cagcaagaag cagagggttg ctcccagggc
     3841 aagaggcaaa tcatccgaac ccgtggtcat catgaagaga agtttgagga cttctgcaaa
     3901 aagaattgaa cctgcggaag agctgaacag caacgacatg aaaaccaaca aagaggaaca
     3961 caaattacaa gactcggtcc ctgaaaataa gggaatatcc ctgcgctcca gacgccaaaa
     4021 taagactgag gcagaacagc aaataactga ggtctttgta ttagcagaaa gaatagaaat
     4081 aaacagaaat gaaaagaagc ccatgaagac ctccccagag atggacattc agaatccaga
     4141 tgatggagcc cggaaaccca tacctagaga caaagtcact gagaacaaaa ggtgcttgag
     4201 gtctgctaga cagaatgaga gctcccagcc taaggtggca gaggagagcg gagggcagaa
     4261 gagtgcgaag gttctcatgc agaatcagaa agggaaagga gaagcaggaa attcagactc
     4321 catgtgcctg agatcaagaa agacaaaaag ccagcctgca gcaagcactt tggagagcaa
     4381 atctgtgcag agagtaacgc ggagtgtcaa gaggtgtgca gaaaatccaa agaaggctga
     4441 ggacaatgtg tgtgtcaaga aaataagaac cagaagtcat agggacagtg aagatatttg
     4501 ggatcctatg gtgagcaagg gcgaggagga taacatggcc tctctcccag cgacacatga
     4561 gttacacatc tttggctcca tcaacggtgt ggactttgac atggtgggtc agggcaccgg
     4621 caatccaaat gatggttatg aggagttaaa cctgaagtcc accaagggtg acctccagtt
     4681 ctccccctgg attctggtcc ctcatatcgg gtatggcttc catcagtacc tgccctaccc
     4741 tgacgggatg tcgcctttcc aggccgccat ggtagatggc tccggatacc aagtccatcg
     4801 cacaatgcag tttgaagatg gtgcctccct tactgttaac taccgctaca cctacgaggg
     4861 aagccacatc aaaggagagg cccaggtgaa ggggactggt ttccctgctg acggtcctgt
     4921 gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg aagaagactt accccaacga
     4981 caaaaccatc atcagtacct ttaagtggag ttacaccact ggaaatggca agcgctaccg
     5041 gagcactgcg cggaccacct acacctttgc caagccaatg gcggctaact atctgaagaa
     5101 ccagccgatg tacgtgttcc gtaagacgga gctcaagcac tccaagaccg agctcaactt
     5161 caaggagtgg caaaaggcct ttaccgatgt gatgggcatg gacgagctgt acaagtaagc
     5221 ggccgcatag ataactgatc cagtgtgctg gaattaattc gctgtctgcg agggccagct
     5281 gttggggtga gtactccctc tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt
     5341 ccaaaaacga ggaggatttg atattcacct ggcccgcggt gatgcctttg agggtggccg
     5401 cgtccatctg gtcagaaaag acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg
     5461 agatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
     5521 ccactcccag gtccaactgc aggtcgagca tgcatctagg gcggccaatt ccgcccctct
     5581 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     5641 gtctatatgt gattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     5701 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     5761 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     5821 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     5881 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     5941 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     6001 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     6061 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     6121 gttttccttt gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca
     6181 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     6241 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     6301 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     6361 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     6421 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     6481 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6541 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     6601 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     6661 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6721 acgtcgagtg cccgaaggac cgcgcgacct ggtgcatgac ccgcaagccc ggtgcctgac
     6781 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat ggctccgacc
     6841 gaagccgacc cgggcggccc cgccgacccc gcacccgccc ccgaggccca ccgactctag
     6901 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     6961 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     7021 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     7081 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     7141 tatggcttct gaggcggaaa gaaccagctg gggctcgagt gcattctagt tgtggtttgt
     7201 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     7261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7321 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7861 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8101 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8281 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9421 ctgacgtcga cggatcggga gatctcccga tcccctatgg tcgactctca gtacaatctg
     9481 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     9541 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     9601 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     9661 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     9721 cccatatatg gagttc
//
