LOCUS       sequence-310009-        5317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acute increase in MPA when used with FRB-FKBP dimerizaton system 
            upon addition with Rapamycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5317)
  AUTHORS   Bisaria A, Hayer A, Garbett D, Cohen D, Meyer T
  TITLE     Membrane-proximal F-actin restricts local membrane protrusions and 
            directs cell migration.
  JOURNAL   Science. 2020 Jun 12;368(6496):1205-1210. doi: 
            10.1126/science.aay7794.
  PUBMED    32527825
REFERENCE   2  (bases 1 to 5317)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2020-06-12- 12"; volume: "368"; 
            issue: "6496"; pages: "1205-1210"
COMMENT     Sequence Label: C1-mCitrine-FKBP-EZRabd(t567D)
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     394..413
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(712..731)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    756..803
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1132..1720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1621..1640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1895..2198
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2199..2402
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2353..2373
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2462..2464,2465..2467,2468..3178)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2507..2528)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2768..2787)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3115..3136
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3257..3577
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3939..4060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3976..3995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4030..4049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4067..4522)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4204..4225)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4416..4435
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4549..4653
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4655..5012
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4655..4846
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4678..4698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4863..4998
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5047..5317,1..524)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(5047..5317,1..524)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5101..5120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
       61 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
      121 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
      181 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
      241 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
      301 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
      361 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
      421 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
      481 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
      541 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
      601 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
      661 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
      721 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
      781 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
      841 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
      901 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
      961 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     1021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1741 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1801 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     1861 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1921 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1981 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2041 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2101 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2161 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2221 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2281 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2341 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2401 ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct
     2461 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2521 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2581 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2641 cctcgtgacc accttcggct acggcctgat gtgcttcgcc cgctaccccg accacatgaa
     2701 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2761 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2821 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2881 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2941 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     3001 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3061 ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     3121 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagag
     3181 atctcgagct caagcttcga ccggtggtgg atccggagga accagtgctg gcgggagcgc
     3241 tggtggaagt gcggggggag tccaggtgga aactataagt ccgggtgacg gccggacctt
     3301 tccaaaacgg gggcagactt gtgttgttca ctatactggg atgttggaag atgggaaaaa
     3361 attcgattcc agtcgagacc ggaataaacc gttcaaattc atgttgggta aacaggaagt
     3421 aattcgagga tgggaagagg gagtggctca gatgtctgta ggccaacgag ctaaacttac
     3481 gatctcccca gattacgcct acggagctac tggccacccg ggtattatcc cccctcacgc
     3541 gacgctcgtc tttgacgttg aactccttaa actcgaatct ggtgggacaa gcgcgggtgg
     3601 aagcgctgga ggttctgctg ggggctcagc tggtggatca gctggaggta gcggctccgc
     3661 agggggttct gcaggaggca gcaccagcgc tggcggatca gcaggtggca tgatccacaa
     3721 cgagaacatg aggcaaggcc gggacaagta caaggatctg cggcagatcc ggcagggcaa
     3781 caccaagcag cgcatcgacg agttcgaggc cctgtaagcg gccgcgactc tagatcataa
     3841 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3901 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3961 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4021 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     4081 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4141 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4201 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4261 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     4321 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     4381 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     4441 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4501 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4621 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4681 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4741 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4801 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4861 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4921 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4981 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     5041 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5101 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5161 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5221 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5281 gctgtgctcg acgttgtcac tgaagcggga agggact
//
