LOCUS       sequence-269442-        5682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golgi-resident FRB for chemically induced dimerization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5682)
  AUTHORS   Zewe JP, Miller AM, Sangappa S, Wills RC, Goulden BD, Hammond GRV
  TITLE     Probing the subcellular distribution of phosphatidylinositol reveals
            a surprising lack at the plasma membrane.
  JOURNAL   J Cell Biol. 2020 Mar 2;219(3). pii: 133808. doi: 
            10.1083/jcb.201906127.
  PUBMED    32211893
REFERENCE   2  (bases 1 to 5682)
  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"; journalName: "J Cell 
            Biol."; date: "2020-03-2"; volume: "219(3). pii: 133808. doi"; 
            pages: " 10.1083/jcb.201906127"
COMMENT     Sequence Label: piRFP-FRB-Giantin
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1560
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     CDS             1609..1887
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     polyA_signal    2470..2591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2507..2526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2561..2580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2598..3053)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2735..2756)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2947..2966
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3080..3184
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3186..3543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3186..3377
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3209..3229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3394..3529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3578..4372
                     /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             3578..4372
                     /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(3632..3651)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4242..4261
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4560..4579)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4604..4651
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4980..5568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5469..5488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggcgga aggatccgtc gccaggcagc ctgacctctt gacctgcgac
      661 gatgagccga tccatatccc cggtgccatc caaccgcatg gactgctgct cgccctcgcc
      721 gccgacatga cgatcgttgc cggcagcgac aaccttcccg aactcaccgg actggcgatc
      781 ggcgccctga tcggccgctc tgcggccgat gtcttcgact cggagacgca caaccgtctg
      841 acgatcgcct tggccgagcc cggggcggcc gtcggagcac cgatcactgt cggcttcacg
      901 atgcgaaagg acgcaggctt catcggctcc tggcatcgcc atgatcagct catcttcctc
      961 gagctcgagc ctccccagcg ggacgtcgcc gagccgcagg cgttcttccg ccgcaccaac
     1021 agcgccatcc gccgcctgca ggccgccgaa accttggaaa gcgcctgcgc cgccgcggcg
     1081 caagaggtgc ggaagattac cggcttcgat cgggtgatga tctatcgctt cgcctccgac
     1141 ttcagcggcg aagtgatcgc agaggatcgg tgcgccgagg tcgagtcaaa actaggcctg
     1201 cactatcctg cctcaaccgt gccggcgcag gcccgtcggc tctataccat caacccggta
     1261 cggatcattc ccgatatcaa ttatcggccg gtgccggtca ccccagacct caatccggtc
     1321 accgggcggc cgattgatct tagcttcgcc atcctgcgca gcgtctcgcc cgtccatctg
     1381 gaattcatgc gcaacatagg catgcacggc acgatgtcga tctcgatttt gcgcggcgag
     1441 cgactgtggg gattgatcgt ttgccatcac cgaacgccgt actacgtcga tctcgatggc
     1501 cgccaagcct gcgagctagt cgcccaggtt ctggcctggc agatcggcgt gatggaagag
     1561 tccggactca gatctcgagc tggaggtgct ggtgctatcc tatctagaat cctctggcat
     1621 gagatgtggc atgaaggcct ggaagaggca tctcgtttgt actttgggga aaggaacgtg
     1681 aaaggcatgt ttgaggtgct ggagcccttg catgctatga tggaacgggg cccccagact
     1741 ctgaaggaaa catcctttaa tcaggcctat ggtcgagatt taatggaggc ccaagagtgg
     1801 tgcaggaagt acatgaaatc agggaatgtc aaggacctca cccaagcctg ggacctctat
     1861 tatcatgtgt tccgacgaat ctcaaagggt ggtagtgctg gtggtagtgc tgaaccgcag
     1921 caaagctttt ctgaagctca gcagcagcta tgcaacacca gacaggaagt gaatgaatta
     1981 aggaagctgc tggaagaaga acgagaccaa agagtggctg ctgagaatgc tctctctgtg
     2041 gccgaggagc agatcagacg gttagagcac agtgaatggg actcttcccg gactcctatc
     2101 attggctcct gtggcactca ggagcaggca ctgttaatag atcttacaag caacagttgt
     2161 cgaaggaccc ggagtggcgt tggatggaag cgagtcctgc gttcactctg tcattcacgg
     2221 acccgagtgc cacttctagc agccatctac tttctaatga ttcatgtcct gctcattctg
     2281 tgttttacgg gccatctata gcaagcttcg aattctgcag tcgacggtac cgcgggcccg
     2341 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2401 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2461 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2521 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2581 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2641 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2701 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2761 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2821 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2881 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2941 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3001 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3061 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3121 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3181 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3301 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3361 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3421 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3481 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3541 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3601 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3661 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3721 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3781 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3841 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3901 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3961 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4021 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4081 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4141 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4201 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4261 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4321 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4381 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4441 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4501 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4561 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4621 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4681 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4741 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4801 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4861 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4921 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4981 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5041 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5101 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5161 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5221 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5281 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5341 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5401 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5461 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5521 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5581 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5641 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
