LOCUS       pcDNA5/FRT/TO_[Cox8a]x4_SNAP-tag2_T2A_EGFP        6793 bp    DNA     circular UNA 03-SEP-2024
DEFINITION  pcDNA5/FRT/TO_[Cox8a]x4_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"
     primer_bind     complement(1024..1057)
                     /standard_name="jh3918"
     primer_bind     1034..1061
                     /standard_name="jh3915"
     CDS             1046..1402
                     /codon_start=1
                     /product="kozac-Cox8a-Cox8a"
                     /note="kozac-Cox8a-Cox8a"
                     /standard_name="Cox8ax4 (Mito targeting sequence)"
     primer_bind     complement(1384..1414)
                     /Sequence="GCAGTCTTTATCGCTTCCACCAGAACCTCCG"
                     /%GC="54.8"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="jh3919"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:fls-g6zazci"">jh3919</a>"
                     /label="jh3919"
     primer_bind     1391..1432
                     /Sequence="TCTGGTGGAAGCGATAAAGACTGCGAAATGAAACGCACTACC"
                     /%GC="47.6"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="jh3920"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:flt-g6zazci"">jh3920</a>"
                     /label="jh3920"
     CDS             <1403..1915
                     /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           1433..1435
                     /created_by="Stefanie"
                     /label="L12Y"
     motif           1457..1459
                     /created_by="Stefanie"
                     /label="E20L"
     motif           1487..1489
                     /created_by="Stefanie"
                     /label="E30R"
     motif           1493..1495
                     /created_by="Stefanie"
                     /label="I32Y"
     motif           1499..1501
                     /standard_name="L34V"
     motif           1505..1507
                     /created_by="Stefanie"
                     /label="K36N"
     motif           1508..1531
                     /created_by="Stefanie"
                     /label="8aa loop design 1"
     motif           1715..1717
                     /created_by="Stefanie"
                     /label="H116Q"
     motif           1772..1774
                     /created_by="Stefanie"
                     /label="S135R"
     motif           1826..1828
                     /created_by="Stefanie"
                     /label="L153G"
     motif           1850..1852
                     /created_by="Stefanie"
                     /label="G161P"
     motif           1856..1858
                     /created_by="Stefanie"
                     /label="A163Y"
     primer_bind     complement(1898..1927)
                     /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     1904..1929
                     /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             1931..1984
                     /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             1985..2698
                     /standard_name="mEGFP"
     primer_bind     complement(2679..2716)
                     /standard_name="Jh3199"
     primer_bind     2693..2722
                     /standard_name="Jh3200"
     motif           2699..2704
                     /standard_name="STOPx2"
     Terminator      2748..2975
                     /standard_name="bGH_PA term"
     misc_feature    3038..3344
                     /standard_name="f1 origin"
                     /note="Geneious type: Origin of Replication"
     Gene            3259..3306
                     /standard_name="FRT"
     Marker          3302..4333
                     /standard_name="hygroB marker"
     Promoter        complement(4640..4714)
                     /standard_name="trc prom"
     misc_feature    complement(4654..4681)
                     /standard_name="lacO reg"
                     /note="Geneious type: Regulatory Sequence"
     Promoter        complement(4685..4714)
                     /standard_name="lac prom"
     misc_feature    complement(5023..5642)
                     /standard_name="pBR322 origin"
                     /note="Geneious type: Origin of Replication"
     Marker          complement(5797..6657)
                     /standard_name="amp marker"
     Promoter        complement(6699..6727)
                     /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 ccaccatgtc ggtcctcacg ccactgttgc tgcgaggttt
     1081 gacaggttcc gcccggcgcc tcccggtgcc acgcgctaaa attcattctt tgtccgtgct
     1141 tacgccgttg ctgttgcggg gattgactgg ttctgctaga cggcttccag ttccacgtgc
     1201 taaaatccat tcgctctctg tccttacgcc actgctgctg cggggcttga caggatctgc
     1261 tcgacgactc ccagtgccgc gcgcaaagat ccattcgttg tccgtcctga cgccactcct
     1321 cctgcggggt ctgacaggtt cggcccgccg acttccagtg ccacgagcga agatccactc
     1381 tctcggaggt tctggtggaa gcgataaaga ctgcgaaatg aaacgcacta cctatgacag
     1441 cccgctgggt aaactgctgc tgagcggttg cgagcagggc ctgcaccgca tctatttcgt
     1501 gggcaatggt cagggcgagc agggtcctcc gggcccggag ccgctgatgc aggctaccgc
     1561 atggctgaac gcatattttc accagccaga ggcgattgaa gagttccctg taccggccct
     1621 gcatcacccg gtatttcagc aggaatcctt tacccgtcag gtactgtgga aactgctgaa
     1681 agtagtgaaa ttcggcgagg tgatctccta ttcccagctg gcggcgctgg ctggcaaccc
     1741 agcggcaact gcggcggtga aaaccgctct gcgtggtaat ccggtgccaa ttctgatccc
     1801 gtgccaccgt gtggttcagg gcgacggcga cgtaggtggc tacgaaggcc ctctgtacgt
     1861 gaaagaatgg ctgctggcgc acgaaggtca ccgtctgggt aaaccaggcc tgggcggccg
     1921 cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     1981 cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2041 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2101 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2161 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2221 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2281 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2341 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2401 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2461 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2521 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2581 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2641 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2701 gtaaccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2761 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2821 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2881 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2941 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3001 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3061 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3121 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3181 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3241 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     3301 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     3361 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     3421 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     3481 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     3541 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     3601 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3661 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     3721 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3781 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     3841 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     3901 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     3961 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     4021 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     4081 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     4141 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     4201 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     4261 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     4321 gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag
     4381 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4441 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4501 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4561 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4621 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4681 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4741 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4801 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4861 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4921 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4981 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5041 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5101 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5161 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5221 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5281 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5341 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5401 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5461 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5521 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5581 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5641 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5701 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5761 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5821 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5881 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5941 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6001 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6061 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6121 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6181 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6241 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6301 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6361 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6421 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6481 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6541 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6601 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6661 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6721 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6781 gccacctgac gtc
//
