LOCUS       sequence-194065-        8346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HIP1R with a GFP-FKBP tag in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8346)
  AUTHORS   Clarke NI, Royle SJ
  TITLE     FerriTag is a new genetically-encoded inducible tag for correlative 
            light-electron microscopy.
  JOURNAL   Nat Commun. 2018 Jul 4;9(1):2604. doi: 10.1038/s41467-018-04993-0.
  PUBMED    29973588
REFERENCE   2  (bases 1 to 8346)
  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: "Nat 
            Commun."; date: "2018-07-4"; volume: "9(1):2604. doi"; pages: " 
            10.1038/s41467-018-04993-0"
COMMENT     Sequence Label: HIP1R-GFP-FKBP
FEATURES             Location/Qualifiers
     source          1..8346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      34..622
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     523..542
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        797..1100
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1101..1304
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1255..1275
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1336..1360
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4571..4595)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4642..4651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4648..4650,4651..4653,4654..5361)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     CDS             join(4648..4650,4651..4653,4654..5361)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(4693..4714)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4954..4973)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5301..5322
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(5410..5412,5413..5736)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    5870..5991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5907..5926)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5961..5980
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5998..6453)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6135..6156)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6347..6366
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6480..6584
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6586..6943
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6586..6777
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6609..6629)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6794..6929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6978..7772
                     /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             6978..7772
                     /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(7032..7051)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7642..7661
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7960..7979)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8004..8051
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
       61 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      121 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      181 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      241 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      301 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      361 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      421 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      481 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
      541 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
      601 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
      661 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
      721 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      781 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      841 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      901 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      961 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     1021 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     1081 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     1141 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     1201 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     1261 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     1321 agatccgcta gcatcacaag tttgtacaaa aaagcaggct taaccatgaa cagcatcaag
     1381 aatgtgccgg cgcgggtgct gagccgcagg ccgggccaca gcctagaggc cgagcgcgag
     1441 cagttcgaca agacgcaggc catcagtatc agcaaagcca tcaacagcca ggaggcccca
     1501 gtgaaggaga agcatgcccg gcgtatcatc ctgggcacgc atcatgagaa gggagccttt
     1561 accttctggt cctatgccat cggcctgccg ctgtccagca gctccatcct cagctggaag
     1621 ttctgtcacg tccttcacaa ggtcctccgg gacggacacc ccaacgtcct gcatgactat
     1681 cagcggtacc ggagcaacat acgtgagatc ggtgacttgt ggggccacct tcgtgaccag
     1741 tatggacacc tggtgaatat ctataccaaa ctgttgctga ctaagatctc cttccacctt
     1801 aagcatcccc agtttcctgc aggcctggag gtaacagatg aggtgttgga gaaggcggcg
     1861 ggaactgatg tcaacaacat ttttcagctt accgtggaga tgtttgacta catggactgt
     1921 gaactgaagc tttctgagtc agttttccgg cagctcaaca cggccatcgc agtgtcccag
     1981 atgtcttctg gccagtgtcg cctagcgccg ctcatccagg tcattcagga ctgcagccac
     2041 ctgtaccact acacagtgaa gctcatgttt aagctgcact cctgtctccc ggcagacacc
     2101 ctgcaaggcc acagggatcg gttccacgag cagtttcaca gcctcaaaaa cttcttccgc
     2161 cgagcttcag acatgctgta cttcaagagg ctcatccaga tcccgcggct gcctgaggga
     2221 ccccccaatt tcctgcgggc ttcagccctg gctgagcaca tcaagccggt ggtggtgatt
     2281 cccgaggagg ccccagagga agaggagcct gagaacctaa ttgaaatcag cagtgcaccc
     2341 cctgctgggg agccagtggt ggtggctgac ctctttgatc agacctttgg accccccaat
     2401 ggctccatga aggatgacag ggacctccaa atcgagaact tgaagagaga ggtggagacc
     2461 ctccgtgctg agctggagaa gattaagatg gaggcacagc ggtacatctc ccagctgaag
     2521 ggccaggtga atggcctgga ggcagagctg gaggagcagc gcaagcagaa gcagaaggcc
     2581 ctggtggaca acgagcagct gcgccacgag ctggcccagc tcaaggccct gcagctggag
     2641 ggcgcccgca accagggcct tcgagaggaa gcagagagga aggccagtgc cacggaggca
     2701 cgctacagca agctgaagga gaaacacagc gaactcatta acacccacgc cgagctgctc
     2761 aggaagaacg cagacacggc caagcagctg acagtgacac agcagagcca ggaggaggtg
     2821 gcacgggtaa aggaacagct ggccttccag atggagcaag cgaagcgtga gtctgagatg
     2881 aagatggaag agcagagcga ccagttggag aagctcaaga gggagctggc ggccagggca
     2941 ggagagctgg cccgtgcgca ggaggccctg agccgcacag aacagagtgg gtcagagctg
     3001 agctcacggc tggacacact gaatgcggag aaggaagctc tgagtggagt cgttcggcag
     3061 cgtgaggcag agctgctggc cgctcagagc ctggtgcggg agaaggagga ggcacttagc
     3121 caagagcagc agcggagctc ccaggagaag ggcgagctac gggggcagct ggcagaaaag
     3181 gagtctcagg agcaggggct tcggcagaag ctgctggatg agcagttggc ggtgttgcga
     3241 agtgcagccg ccgaggcaga ggccatccta caggatgcag tgagcaagct ggacgacccc
     3301 ctgcacctcc gctgcaccag ctccccagac tacttggtga gccgggctca ggcagccctg
     3361 gacagcgtga gcggcctgga gcagggccac acccagtacc tggcttcctc tgaagatgct
     3421 tctgccctgg tggcagcgct gacccgcttc tcccatttgg ctgcggacac cattgtcaat
     3481 ggtgccgcca cctcccacct ggcccccacc gaccccgccg accgcctgat ggacacatgc
     3541 agggagtgtg gagcccgggc tctggagctg gtgggacagc tgcaagacca gacagtgcta
     3601 ccgagggctc agcccagcct gatgcgggcc cccctgcagg gcattctgca gttgggccag
     3661 gacttgaagc ctaagagcct ggatgtacgg caagaggagc taggggccat ggtggacaag
     3721 gagatggcgg ccacctcggc agccattgag gacgctgtgc ggaggatcga ggacatgatg
     3781 agccaggccc gccacgagag ctcgggcgtg aaactggagg tgaatgagag gatcctcaac
     3841 tcctgcacag acctgatgaa ggctatccgg ctcctggtga tgacctccac cagcctgcag
     3901 aaggaaattg tggagagcgg caggggggca gcaacgcagc aggaatttta tgccaagaat
     3961 tcacggtgga ctgaaggcct catctcagcc tctaaggcag tgggctgggg agccacacag
     4021 ctggtggagt cagctgacaa ggttgtgctt cacatgggca aatacgagga actcatcgtc
     4081 tgctcccatg agattgcggc cagcacggcc cagctggtgg cagcctcgaa ggtgaaagcc
     4141 aacaagaaca gtccccactt gagccgcctg caggaatgtt cccgcactgt caacgagagg
     4201 gctgcctacg tcgtggcctc caccaaatct ggccaggagc agattgagga cagagacacc
     4261 atggatttct ctggcctgtc cctcatcaag ttgaagaagc aggagatgga gacacaggtg
     4321 cgagtcttgg agctggagaa gacactagag gcagagcgtg tccggctcgg ggagcttcgg
     4381 aaacagcact atgtactggc tggggggatg ggaacaccta gcgaagaaga acccagcaga
     4441 cccagcccag ctccccgaag tggggccact aagaagccac cgctggccca gaaacccagc
     4501 atagccccca ggacagacaa ccagctcgac aaaaaggatg gtgtctaccc agctcaactt
     4561 gtgaactacg acccagcttt cttgtacaaa gtggtgggac atggtacagg atcaactggt
     4621 agtggaagta gtataccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     4681 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4741 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     4801 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     4861 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4921 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     4981 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     5041 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     5101 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     5161 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     5221 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     5281 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     5341 ctcggcatgg acgagctgta ccgaatcgat gtgaatgggg gccctgaacc cgccaagaac
     5401 ctgtacacca tgggagtgca ggtggaaacc atctccccag gagacgggcg caccttcccc
     5461 aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaattt
     5521 gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc
     5581 cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata
     5641 tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact
     5701 ctcgtcttcg atgtggagct tctaaaactg gaataaggta caagtaaagc ggccgcgact
     5761 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     5821 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     5881 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5941 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     6001 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6061 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     6121 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6181 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6241 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6301 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6361 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6421 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     6481 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6541 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     6601 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     6661 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     6721 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     6781 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6841 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6901 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     6961 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     7021 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     7081 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     7141 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     7201 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     7261 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     7321 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     7381 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     7441 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     7501 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     7561 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     7621 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     7681 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     7741 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     7801 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7861 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7921 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     7981 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     8041 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     8101 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     8161 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     8221 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8281 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8341 ctgagc
//
