LOCUS       Pβactin-FKBP-mSc        8795 bp    DNA     circular UNA 01-SEP-2022
DEFINITION  Gibson Assembly of 1_RhoA-BS_PCR#1 into Digestion of pCAG-AcGFP-CI
            (modified) - Fragment 2.
ACCESSION   urn.local...9h-f94o2g8
VERSION     urn.local...9h-f94o2g8
KEYWORDS    .
SOURCE      3727673.30 Da
  ORGANISM  3727673.30 Da.
FEATURES             Location/Qualifiers
     polya_signal    43..271
                     /type="polyA_signal"
                     /Transferred_From="pCAG-mCherry-LFC"
                     /Transferred_Similarity="100.00%"
                     /label="SV40 early polyA signal"
     misc_feature    complement(182..211)
                     /Mismatches=0
                     /%GC="26.7"
                     /Hairpin_Tm="40.5"
                     /Self_Dimer_Tm="None"
                     /Tm="60.1"
                     /annotation_group="164504135370387a40f0291-affb-4892-95c1-
                     432e81482e77"
                     /Sequence="TTTGTGAAATTTGTGATGCTATTGCTTTAT"
                     /created_by="primer3"
                     /label="pEGFP-C RP "
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    complement(187..206)
                     /%GC="35.0"
                     /Mismatches=0
                     /Hairpin_Tm="40.2"
                     /Self_Dimer_Tm="None"
                     /Tm="51.5"
                     /annotation_group="164503933580982cc3dfb5a-52c3-45f9-8380-
                     5bcfb54c962e;
                     164504037061625b1c1c106-cb12-419e-989d-b0a3a0bfa767"
                     /Sequence="GAAATTTGTGATGCTATTGC"
                     /created_by="primer3"
                     /label="SV40pA-R  ; SV40pA-R  "
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     241..260
                     /Mismatches=0
                     /%GC="45.0"
                     /Hairpin_Tm="48.9"
                     /Self_Dimer_Tm="19.2"
                     /Tm="54.8"
                     /annotation_group="164504431238753d69d49bb-2d64-4d34-a2c0-
                     513bd4ded39c"
                     /Sequence="GTGGTTTGTCCAAACTCATC"
                     /created_by="primer3"
                     /label="AcGFP Reverse "
     misc_feature    complement(298..314)
                     /Mismatches=0
                     /%GC="47.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="49.1"
                     /annotation_group="1645042358568559838f31d-246c-409c-98c1-
                     a066ae9a2406"
                     /Sequence="CAGGAAACAGCTATGAC"
                     /created_by="primer3"
                     /label="M13 Reverse  "
                     /note="Geneious type: primer_bind_reverse"
     cds             complement(1459..2319)
                     /type="CDS"
                     /Transferred_From="pCAG-mCherry-LFC"
                     /Transferred_Similarity="100.00%"
                     /label="AmpR"
     misc_feature    complement(2549..2567)
                     /Mismatches=0
                     /%GC="57.9"
                     /Hairpin_Tm="33.5"
                     /Self_Dimer_Tm="None"
                     /Tm="58.2"
                     /annotation_group="5521807483510197a5a7ae2-fccd-46c2-99d1-
                     db002ec4044c"
                     /Sequence="GGAGCTGCATGTGTCAGAG"
                     /created_by="primer3"
                     /label="pGEX-3"
                     /note="Geneious type: primer_bind_reverse"
     eukaryotic      2860..4992
                     /promoter_type="eukaryotic"
                     /type="eukaryotic"
                     /Transferred_From="pCAG-mCherry-LFC"
                     /Transferred_Similarity="100.00%"
                     /label="chicken Beta-actin"
     primer_bind     2897..2914
                     /Mismatches=0
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="2.2"
                     /Tm="56.8"
                     /annotation_group="552180642196370b55eb636-6574-40c9-9052-
                     e92d372c1455"
                     /Sequence="TGTAAAACGACGGCCAGT"
                     /created_by="primer3"
                     /label="M13-FP"
     primer_bind     2897..2914
                     /Mismatches=0
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="2.2"
                     /Tm="56.8"
                     /annotation_group="1645043335611165632130e-69c0-4737-bfec-
                     4471484d7a7b"
                     /Sequence="TGTAAAACGACGGCCAGT"
                     /created_by="primer3"
                     /label="M13 (-21) Forward  "
     intron          5184..5993
                     /type="intron"
                     /Transferred_From="pCAG-mCherry-LFC"
                     /Transferred_Similarity="100.00%"
                     /label="intron of chicken Beta-actin"
     primer_bind     5934..5952
                     /Mismatches=0
                     /%GC="63.2"
                     /Hairpin_Tm="56.5"
                     /Self_Dimer_Tm="11.4"
                     /Tm="62.0"
                     /annotation_group="164504503605844a7e03baa-1479-4f7a-959e-
                     16f32481b2a2"
                     /Sequence="GTTCGGCTTCTGGCGTGTG"
                     /created_by="primer3"
                     /label="pCAG_AcGFP_fwd1     "
     source          <5986..>6011
                     /organism="pEGFP-C1"
                     /mol_type="other DNA"
                     /Transferred_From="pCAG-Cdc422G, pCAG-DORARhoA(Q63L),
                     pCAG-DORARhoA(wt), pCAG-RaichuCRRhoA, pCAG-mCherry-GEFH1,
                     pCAG-mCherry-LFC, pCAG-mClover, pCAG-mClover-mRuby2,
                     pCAG-mRuby2, pCAG-mRuby2-GEFH1, pCAG-mRuby2-LFC,
                     pCAG-mRuby2-p190RhoGAP, pCAG-mTFP, pCAG-mTFP-GEFH1,
                     pCAG-mTFP-LFC, pCAG-mTFP-mVenus, pCAG-mVenus,
                     pCAG-meGFP-mCherry, pCAG-p190RhoGAP,
                     pCAG_mCherry-p190RhoGAP"
                     /Transferred_Similarity="100.00%"
                     /label="source pEGFP-C1"
     misc_feature    6011
                     /created_by="schelskim"
                     /label="lastNtBeforeIntegration"
     CDS             6012..6332
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVE
                     LLKLE"
                     /label="FKBP"
     misc_feature    6333..6353
                     /created_by="schelskim"
                     /modified_by="schelskim"
                     /label="HSLink1"
     CDS             6354..7049
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved
                      from a synthetic template (Bindels et al., 2016)"
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQT
                     AKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFED
                     GGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKG
                     DIKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERS
                     EGRHSTGGMDELYK"
                     /modified_by="schelskim"
                     /label="mScarlet"
     misc_feature    7050..7070
                     /created_by="schelskim"
                     /label="HSLink2"
     CDS             7071..8717
                     /created_by="schelskim"
                     /modified_by="schelskim"
                     /label="KIFC1Motor"
     CDS             8718..8762
                     /created_by="schelskim"
                     /modified_by="schelskim"
                     /label="HAtag"
ORIGIN      
        1 acggtaccgc gggcccggga tccaccggat ctagataagt aatgatcata atcagccata
       61 tcacatctgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
      121 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
      181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaattcg taatcatgtc
      301 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      361 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      421 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1141 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2461 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2521 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2581 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2641 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2701 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     2761 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     2821 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     2881 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ttgggatctt tgcattggcc
     2941 cacggctctc aggatgggga tgctcccctt cagcacccgg ttccccttgg aaactgatgg
     3001 tcctggctct gtggcatggc agtggcactg tgaggagccc cctaccagca gcacacagtg
     3061 gggttggcac tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac
     3121 ccgaattgcc ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc
     3181 tgccttctgg tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt
     3241 cccattgccg ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg
     3301 cccttctttg acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac
     3361 gttaattaat tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg
     3421 gggctgatac agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca
     3481 cggaggtccc catgtgctgc agtgtcaccg cctccgagga gacagtattg tccctgcggt
     3541 gtccctgcag ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag
     3601 ccccgatgca acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac
     3661 acagggctgg gtcccgctcc cctaatatct acagtgcttc ttgcatggcc ccttaatcaa
     3721 tgcagttaat cagcatgcgc tcatgcaccg ctctggagct gcaaagcccc tcgcagcgct
     3781 gctcaccaac accgcgcacc gccccggccc agcctgcagc acgcgctgca aacaggaaag
     3841 aaacaaaata ttgcccaaat gtaggcaaag gcattcggct gccttgacct ccgccgggcc
     3901 gggccctgcc tgactcagct ccttactcag cgctcgcttc ctccctccgg ctgccaccgc
     3961 cgcagcgcac accctgacaa agagtggccc ttaacgggct ctgaggtgca cccagcagtg
     4021 cactcagcag tccaagggcc ggcctggagg tttgcaccgc tacgtgctga cattagcatt
     4081 gaacttggcc ctgggtagtg ctgcaggccg ggcggggtgg gtgtagagag tgcagcgcgc
     4141 gttgcacccg gtgccccttc ccctcccttg catcccagca ggctgcaccc cagcaccagg
     4201 cccgtgcatg catgctcctg gtgttattgc agcctggtgc atgcatgcgt cttgatggtg
     4261 cagcgctgtg catgcatcct ccttggtgtg tagcagtcta gtgcatgcat acccctcggt
     4321 gttattgctg ctctgtgcac gcacgctcat tgtatcactt catcccagtg catgcactca
     4381 cactggagcg attgctgctc ggtgcacgca cactcattgt atcactgcag ctcagtggct
     4441 gcacgcacac cggtgttatt gctgctcggt gcgtgcatgc acatcagtgt cgctgcagct
     4501 cagtgcatgc acgctcattg cccatcgcta tccctgcctc tcctgctggc gctccccggg
     4561 aggtgacttc aaggggaccg caggaccacc tcgggggtgg ggggagggct gcacacgcgg
     4621 accccgctcc ccctccccaa caaagcactg tggaatcaaa aaggggggag gggggatgga
     4681 ggggcgcgtc acacccccgc cccacaccct cacctcgagg tgagccccac gttctgcttc
     4741 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     4801 ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     4861 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     4921 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     4981 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     5041 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     5101 cccttctcct ccgggctgta attagcgctt ggtttaatga cggctcgttt cttttctgtg
     5161 gctgcgtgaa agccttaaag ggctccggga gggccctttg tgcggggggg agcggctcgg
     5221 ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg
     5281 gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga
     5341 gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg
     5401 tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac
     5461 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
     5521 cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg
     5581 ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc
     5641 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
     5701 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
     5761 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
     5821 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctcca tctccagcct
     5881 cggggctgtc cgcaggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     5941 ttctggcgtg tgaccggcgg ggtttatatc ttcccttctc tgttcctccg cagcccccaa
     6001 gctagctcga gggagtgcag gtggaaacca tctccccagg agacgggcgc accttcccca
     6061 agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaatttg
     6121 attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc
     6181 gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat
     6241 ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca catgccactc
     6301 tcgtcttcga tgtggagctt ctaaaactgg aaggaagttc aggaggttct agtatggtga
     6361 gcaagggcga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg gagggctcca
     6421 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     6481 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg gacatcctgt
     6541 cccctcagtt catgtacggc tccagggcct tcatcaagca ccccgccgac atccccgact
     6601 actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     6661 gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg atctacaagg
     6721 tgaagctccg cggcaccaac ttccctcctg acggccccgt aatgcagaag aagacaatgg
     6781 gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag ggcgacatta
     6841 agatggccct gcgcctgaag gacggcggcc gctacctggc ggacttcaag accacctaca
     6901 aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag ttggacatca
     6961 cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag ggccgccact
     7021 ccaccggcgg catggacgag ctgtacaagt ctggtggatc aagtggtgga atggcaggag
     7081 ggaagaaacc cagcaaacgt ccagcctggg acttaaaggg tcagttatgt gacctaaatg
     7141 cagaactaaa acggtgccgt gagaggactc aaacgttgga ccaagagaac cagcagcttc
     7201 aggaccagct cagagatgcc cagcagcagg tcaaggccct ggggacagag cgcacaacac
     7261 tggaggggca tttagccaag gtacaggccc aggctgagca gggccaacag gagctgaaga
     7321 acttgcgtgc ttgtgtcctg gagctggaag agcggctgag cacgcaggag ggcttggtgc
     7381 aagagcttca gaaaaaacag gtggaattgc aggaagaacg gaggggactg atgtcccaac
     7441 tagaggagaa ggagaggagg ctgcagacat cagaagcagc cctgtcaagc agccaagcag
     7501 aggtggcatc tctgcggcag gagactgtgg cccaggcagc cttactgact gagcgggaag
     7561 aacgtcttca tgggctagaa atggagcgcc ggcgactgca caaccagctg caggaactca
     7621 agggcaacat ccgtgtattc tgccgggtcc gccctgtcct gccgggggag cccactccac
     7681 cccctggcct cctcctgttt ccctctggcc ctggtgggcc ctctgatcct ccaacccgcc
     7741 ttagcctctc ccggtctgac gagcggcgtg ggaccctgag tggggcacca gctcccccaa
     7801 ctcgccatga tttttccttt gaccgggtat tcccaccagg aagtggacag gatgaagtgt
     7861 ttgaagagat tgccatgctt gtccagtcag ccctggatgg ctatccagta tgcatctttg
     7921 cctatggcca gacaggcagt ggcaagacct tcacaatgga gggtgggcct gggggagacc
     7981 cccagttgga ggggctgatc cctcgggccc tgcggcacct cttctctgtg gctcaggagc
     8041 tgagtggtca gggctggacc tacagctttg tagcaagcta cgtagagatc tacaatgaga
     8101 ctgtccggga cctgctggcc actggaaccc ggaagggtca agggggcgag tgtgagattc
     8161 gccgtgcagg gccagggagt gaggagctca ctgtcaccaa tgctcgatat gtccctgtct
     8221 cctgtgagaa agaagtggac gccctgcttc atctggcccg ccagaatcgg gctgtggccc
     8281 gcacagccca gaatgaacgg tcatcacgca gccacagtgt attccagcta cagatttctg
     8341 gggagcactc cagccgaggc ctgcagtgtg gggcccccct cagtcttgtg gacctggccg
     8401 ggagtgagcg acttgacccc ggcttagccc tcggccccgg ggagcgggaa cgccttcggg
     8461 aaacacaggc cattaacagc agcctgtcca cgctggggct ggttatcatg gccctgagca
     8521 acaaggagtc ccacgtgcct taccggaaca gcaaactgac ctacctgctg cagaactctc
     8581 tgggtggtag tgctaagatg ctcatgtttg tgaacatttc tccactggaa gagaacgtct
     8641 ccgagtccct caactctcta cgctttgcct ccaaggtgaa ccagtgtgtt attggtactg
     8701 ctcaggccaa caggaagctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     8761 cctaattaat ctagaagctt cgaattctgc agtcg
//
