LOCUS       pcDNA5/FRT/TO_TOMM20-SNAP-tag2_T2A_EGFP        6898 bp    DNA     circular UNA 03-SEP-2024
DEFINITION  pcDNA5/FRT/TO_TOMM20-SNAP-tag2_T2A_EGFP.
ACCESSION   urn.local...m-b5ympd4
VERSION     urn.local...m-b5ympd4
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     Gene            252..973
                     /standard_name="5_LTR"
     Promoter        758..840
                     /standard_name="CMV prom"
     misc_feature    769..789
                     /standard_name="CMV forward"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    820..859
                     /standard_name="tetO reg"
                     /note="Geneious type: Regulatory Sequence"
     Promoter        836..955
                     /standard_name="CMV2 prom"
     CDS             1043..1501
                     /standard_name="TOMM20"
     motif           1502..1507
                     /standard_name="linker"
     CDS             <1508..2020
                     /gene="AGTSM"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="Stefanie"
                     /modified_by="Stefanie"
                     /label="SNAP_2.0"
     motif           1538..1540
                     /created_by="Stefanie"
                     /label="L12Y"
     motif           1562..1564
                     /created_by="Stefanie"
                     /label="E20L"
     motif           1592..1594
                     /created_by="Stefanie"
                     /label="E30R"
     motif           1598..1600
                     /created_by="Stefanie"
                     /label="I32Y"
     motif           1604..1606
                     /standard_name="L34V"
     motif           1610..1612
                     /created_by="Stefanie"
                     /label="K36N"
     motif           1613..1636
                     /created_by="Stefanie"
                     /label="8aa loop design 1"
     motif           1820..1822
                     /created_by="Stefanie"
                     /label="H116Q"
     motif           1877..1879
                     /created_by="Stefanie"
                     /label="S135R"
     motif           1931..1933
                     /created_by="Stefanie"
                     /label="L153G"
     motif           1955..1957
                     /created_by="Stefanie"
                     /label="G161P"
     motif           1961..1963
                     /created_by="Stefanie"
                     /label="A163Y"
     primer_bind     complement(2003..2032)
                     /Sequence="ACTGCCGCGGCCGCCCAGGCCTGGTTTACC"
                     /%GC="73.3"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="jh3924"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:fn1-g6zazcs"">jh3924</a>"
                     /label="jh3924"
     primer_bind     2009..2034
                     /Sequence="CCAGGCCTGGGCGGCCGCGGCAGTGG"
                     /%GC="84.6"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="jh3923"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:fn0-g6zazcs"">jh3923</a>"
                     /label="jh3923"
     CDS             2036..2089
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding
                     separatepolypeptides."
                     /standard_name="T2A"
     CDS             2090..2803
                     /standard_name="mEGFP"
     primer_bind     complement(2784..2821)
                     /Sequence="AGACTCGAGCGGTTACTACTTGTACAGCTCGTCCATGC"
                     /%GC="52.6"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="Jh3199"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:ctu-g6zayqu"">Jh3199</a>"
                     /label="Jh3199"
     primer_bind     2798..2827
                     /Sequence="TACAAGTAGTAACCGCTCGAGTCTAGAGGG"
                     /%GC="50.0"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="Jh3200"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:ctv-g6zayqu"">Jh3200</a>"
                     /label="Jh3200"
     motif           2804..>2806
                     /standard_name="STOPx2"
     Terminator      2853..3080
                     /standard_name="bGH_PA term"
     misc_feature    3143..3449
                     /standard_name="f1 origin"
                     /note="Geneious type: Origin of Replication"
     Gene            3364..3411
                     /standard_name="FRT"
     Marker          3407..4438
                     /standard_name="hygroB marker"
     Promoter        complement(4745..4819)
                     /standard_name="trc prom"
     misc_feature    complement(4759..4786)
                     /standard_name="lacO reg"
                     /note="Geneious type: Regulatory Sequence"
     Promoter        complement(4790..4819)
                     /standard_name="lac prom"
     misc_feature    complement(5128..5747)
                     /standard_name="pBR322 origin"
                     /note="Geneious type: Origin of Replication"
     Marker          complement(5902..6762)
                     /standard_name="amp marker"
     Promoter        complement(6804..6832)
                     /standard_name="amp prom"
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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatccagcaa ccatggtggg tcggaacagc gccatcgccg ccggtgtatg
     1081 cggggccctt ttcattgggt actgcatcta cttcgaccgc aaaagacgaa gtgaccccaa
     1141 cttcaagaac aggcttcgag aacgaagaaa gaaacagaag cttgccaagg agagagctgg
     1201 gctttccaag ttacctgacc ttaaagatgc tgaagctgtt cagaagttct tccttgaaga
     1261 aatacagctt ggtgaagagt tactagctca aggtgaatat gagaagggcg tagaccatct
     1321 gacaaatgca attgctgtgt gtggacagcc acagcagtta ctgcaggtct tacagcaaac
     1381 tcttccacca ccagtgttcc agatgcttct gactaagctc ccaacaatta gtcagagaat
     1441 tgtaagtgct cagagcttgg ctgaagatga tgtggaaggc ggtagcgggg atccaccggt
     1501 cggaggtgat aaagactgcg aaatgaaacg cactacctat gacagcccgc tgggtaaact
     1561 gctgctgagc ggttgcgagc agggcctgca ccgcatctat ttcgtgggca atggtcaggg
     1621 cgagcagggt cctccgggcc cggagccgct gatgcaggct accgcatggc tgaacgcata
     1681 ttttcaccag ccagaggcga ttgaagagtt ccctgtaccg gccctgcatc acccggtatt
     1741 tcagcaggaa tcctttaccc gtcaggtact gtggaaactg ctgaaagtag tgaaattcgg
     1801 cgaggtgatc tcctattccc agctggcggc gctggctggc aacccagcgg caactgcggc
     1861 ggtgaaaacc gctctgcgtg gtaatccggt gccaattctg atcccgtgcc accgtgtggt
     1921 tcagggcgac ggcgacgtag gtggctacga aggccctctg tacgtgaaag aatggctgct
     1981 ggcgcacgaa ggtcaccgtc tgggtaaacc aggcctgggc ggccgcggca gtggagaggg
     2041 cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccag tgagcaaggg
     2101 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2161 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2221 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2281 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2341 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2401 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2461 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2521 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2581 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2641 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2701 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2761 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtagtaac cgctcgagtc
     2821 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2881 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2941 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3001 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3061 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3121 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3181 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3241 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3301 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     3361 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     3421 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3481 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3541 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3601 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3661 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3721 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3781 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3841 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     3901 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     3961 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     4021 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4081 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4141 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     4201 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     4261 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4321 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4381 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4441 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4501 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4561 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4681 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4741 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4801 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4861 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4921 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4981 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5041 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5881 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5941 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6001 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6061 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6121 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6181 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6241 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6301 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6361 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6421 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6481 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6541 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6601 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6661 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6721 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6781 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6841 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
