LOCUS       pcDNA5/FRT_HaloTag7-SNAP-tag2-NLS-P2A-NLS-mTurquoise2        7441 bp    DNA     circular UNA 03-SEP-2024
DEFINITION  pcDNA5/FRT-HaloTag7-SNAP-tag2-NLS-P2A-NLS-mTurquoise2.
ACCESSION   urn.local...7y-7vqwj6x
VERSION     urn.local...7y-7vqwj6x
KEYWORDS    .
SOURCE      mamalian
  ORGANISM  mamalian.
FEATURES             Location/Qualifiers
     Promoter        758..846
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="CMV prom"
                     /Transferred_Similarity="82.02%"
                     /label="CMV prom"
     misc_feature    863..880
                     /gene="T7 promoter"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /Transferred_From="T7 promoter"
                     /Transferred_Similarity="100.00%"
     Promoter        863..881
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="T7 prom"
                     /Transferred_Similarity="100.00%"
                     /label="T7 prom"
     misc_feature    902..932
                     /created_by="birgit"
                     /modified_by="birgit"
                     /label="BK_043 forward"
     misc_feature    902..925
                     /created_by="birgit"
                     /modified_by="birgit"
                     /label="24bp overlap"
     misc_signal     911..>916
                     /created_by="User"
                     /modified_by="User"
                     /label="Kozak"
     CDS             917..1810
                     /created_by="User"
                     /label="HaloTag7"
     polylinker      1820..1909
                     /created_by="User"
                     /modified_by="Stefanie"
                     /label="P30"
     CDS             1934..2449
                     /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           1967..1969
                     /created_by="Stefanie"
                     /label="L12Y"
     motif           1991..1993
                     /created_by="Stefanie"
                     /label="E20L"
     motif           2021..2023
                     /created_by="Stefanie"
                     /label="E30R"
     motif           2027..2029
                     /created_by="Stefanie"
                     /label="I32Y"
     motif           2033..2035
                     /standard_name="L34V"
     motif           2039..2041
                     /created_by="Stefanie"
                     /label="K36N"
     motif           2042..2065
                     /created_by="Stefanie"
                     /label="8aa loop design 1"
     motif           2249..2251
                     /created_by="Stefanie"
                     /label="H116Q"
     motif           2306..2308
                     /created_by="Stefanie"
                     /label="S135R"
     motif           2360..2362
                     /created_by="Stefanie"
                     /label="L153G"
     motif           2384..2386
                     /created_by="Stefanie"
                     /label="G161P"
     motif           2390..2392
                     /created_by="Stefanie"
                     /label="A163Y"
     misc_signal     2459..2533
                     /created_by="User"
                     /label="NLS 3x"
     primer_bind     complement(2531..2560)
                     /Sequence="GAAGTTAGTGGCTTGAGGAGAAGCCCGGAG"
                     /%GC="56.7"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="sk696"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:b7-giq2zlr"">sk696</a>"
                     /label="sk696"
     primer_bind     2537..2573
                     /Sequence="GCTTCTCCTCAAGCCACTAACTTCTCCCTGTTGAAAC"
                     /%GC="48.6"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="sk695"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:b6-giq2zlr"">sk695</a>"
                     /label="sk695"
     source          <2549..>2638
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             2549..2605
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding
                     separatepolypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             2618..2638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     CDS             2639..3355
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="User"
                     /label="m_Turquoise_2"
     primer_bind     complement(3335..3367)
                     /Sequence="TCTAGATCATTACTTGTACAGCTCGTCCATGCC"
                     /%GC="45.5"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="sk698"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:b9-giq2zlr"">sk698</a>"
                     /label="sk698"
     primer_bind     3344..3381
                     /Sequence="GAGCTGTACAAGTAATGATCTAGAGGGCCCGTTTAAAC"
                     /%GC="44.7"
                     /note="Derived using Geneious Prime 2023.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="sk697"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:b8-giq2zlr"">sk697</a>"
                     /label="sk697"
     Terminator      3396..3623
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="bGH_PA term"
                     /Transferred_Similarity="100.00%"
                     /label="bGH_PA term"
     Gene            3907..3954
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="FRT"
                     /Transferred_Similarity="100.00%"
                     /label="FRT"
     Marker          3950..4981
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="hygroB marker"
                     /Transferred_Similarity="98.93%"
                     /label="hygroB marker"
     Promoter        complement(5288..5362)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="trc prom"
                     /Transferred_Similarity="82.67%"
                     /label="trc prom"
     misc_feature    complement(5302..5329)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="lacO reg"
                     /Transferred_Similarity="89.29%"
                     /label="lacO reg"
                     /note="Geneious type: Regulatory Sequence"
     Promoter        complement(5333..5362)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="lac prom"
                     /Transferred_Similarity="100.00%"
                     /label="lac prom"
     misc_feature    complement(5671..6290)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="pBR322 origin"
                     /Transferred_Similarity="99.68%"
                     /label="pBR322 origin"
                     /note="Geneious type: Origin of Replication"
     CDS             complement(6445..7305)
                     /created_by="Hiblot"
                     /Transferred_From="AmpR"
                     /Transferred_Similarity="100.00%"
                     /label="AmpR"
     Promoter        complement(7347..7375)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /Transferred_From="amp prom"
                     /Transferred_Similarity="100.00%"
                     /label="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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt accaccatgg gatccgaaat cggtactggc tttccattcg acccccatta
      961 tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc
     1021 tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca
     1081 tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa
     1141 accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc
     1201 cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca
     1261 ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc
     1321 tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac
     1381 caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc
     1441 gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct
     1501 gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga
     1561 gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt
     1621 cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg
     1681 cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct
     1741 gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga
     1801 gatttccggt agcggccgcc ctcctcctcc gccaccgcca ccaccgccgc cgccgccacc
     1861 accacctcct ccgcctcctc caccaccacc gccaccgcct ccaccacctg gcggccgctc
     1921 tcggtctctc gagatggata aagactgcga aatgaaacgc actacctatg acagcccgct
     1981 gggtaaactg ctgctgagcg gttgcgagca gggcctgcac cgcatctatt tcgtgggcaa
     2041 tggtcagggc gagcagggtc ctccgggccc ggagccgctg atgcaggcta ccgcatggct
     2101 gaacgcatat tttcaccagc cagaggcgat tgaagagttc cctgtaccgg ccctgcatca
     2161 cccggtattt cagcaggaat cctttacccg tcaggtactg tggaaactgc tgaaagtagt
     2221 gaaattcggc gaggtgatct cctattccca gctggcggcg ctggctggca acccagcggc
     2281 aactgcggcg gtgaaaaccg ctctgcgtgg taatccggtg ccaattctga tcccgtgcca
     2341 ccgtgtggtt cagggcgacg gcgacgtagg tggctacgaa ggccctctgt acgtgaaaga
     2401 atggctgctg gcgcacgaag gtcaccgtct gggtaaacca ggcctgggcg gagctccgga
     2461 tccaaaaaag aagagaaagg tagatccaaa aaagaagaga aaggtagatc caaaaaagaa
     2521 gagaaaggag ctccgggctt ctcctcaagc cactaacttc tccctgttga aacaagcagg
     2581 ggatgtcgaa gagaatcccg ggccatctag aatggccccc aagaagaaga ggaaggtgat
     2641 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2701 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2761 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2821 cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
     2881 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2941 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3001 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3061 gctggagtac aactacttta gcgacaacgt ctatatcacc gccgacaagc agaagaacgg
     3121 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     3181 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3241 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3301 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg
     3361 atctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3421 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3481 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3541 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3601 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3661 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3721 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3781 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3841 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3901 tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa cttccttggc
     3961 caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     4021 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4081 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4141 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     4201 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4261 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     4321 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     4381 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     4441 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4501 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4561 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4621 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4681 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     4741 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4801 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4861 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     4921 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4981 atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5041 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5101 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5161 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5221 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5281 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5341 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5401 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5461 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5521 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5581 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5641 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5701 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5761 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5821 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5881 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5941 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6001 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6061 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6121 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6181 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6241 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6301 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6361 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6421 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6481 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6541 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6601 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6661 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6721 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6781 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6841 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6901 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6961 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7021 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7081 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7141 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7201 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7261 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7321 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7381 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7441 c
//
