LOCUS       sequence-193574-        6076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a clathrin ""hook"" in mammalian cells to hot-wire 
            endocytosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6076)
  AUTHORS   Wood LA, Larocque G, Clarke NI, Sarkar S, Royle SJ
  TITLE     New tools for ""hot-wiring"" clathrin-mediated endocytosis with 
            temporal and spatial precision.
  JOURNAL   J Cell Biol. 2017 Sep 27. pii: jcb.201702188. doi: 
            10.1083/jcb.201702188.
  PUBMED    28954824
REFERENCE   2  (bases 1 to 6076)
  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. 2017 Sep 27. pii: jcb.201702188. doi: 10.1083/jcb.201702188."
COMMENT     Sequence Label: FKBP-beta2(616-951)-mRuby2
FEATURES             Location/Qualifiers
     source          1..6076
                     /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"
     CDS             join(666..668,669..989)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      1014..1023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2864..2985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2901..2920)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2955..2974
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2992..3447)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3129..3150)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3341..3360
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3474..3578
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3580..3937
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3580..3771
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3603..3623)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3788..3923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3972..4766
                     /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             3972..4766
                     /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(4026..4045)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4636..4655
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4954..4973)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4998..5045
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5374..5962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5863..5882
                     /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 ccggactcag atctcgagag atcgatgtga atgggggccc tgaacccgcc aagaacctgt
      661 acaccatggg agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc ttccccaagc
      721 gcggccagac ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaatttgatt
      781 cctcccggga cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag
      841 gctgggaaga aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc
      901 cagattatgc ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg
      961 tcttcgatgt ggagcttcta aaactggaac ggtaccgcgg gcccgggatc cccgccacca
     1021 tggaacagcc tcaggttatc ccctctcaag gtgatcttct aggggatctt ttaaaccttg
     1081 acctcggtcc cccagtcaat gtgccacagg tgtcctccat gcagatggga gcagtggatc
     1141 tcctaggagg aggactagat agtctgcttg gcagtgacct tggcgggggc attggaggaa
     1201 gtccggcagt gggacaatcc ttcatcccat catcggtccc tgcaaccttt gctccttcac
     1261 ctacacctgc tgtggtcagc agtggactga atgacctgtt tgaactctcc acagggatag
     1321 gcatggcacc tggtggatat gtggctccta aggctgtctg gctacctgca gtaaaggcta
     1381 aaggcttgga gatttccgga acatttactc accgccaagg gcacatctat atggaaatga
     1441 acttcaccaa taaagctctg cagcacatga cagattttgc aatccagttt aacaaaaata
     1501 gctttggtgt catccccagc actcctctgg ccatccatac accactgatg ccaaaccaga
     1561 gcattgatgt ctccctgcct ctcaatacct tgggcccagt catgaagatg gaacctctga
     1621 ataacctcca ggtggctgtg aaaaacaata tcgatgtctt ctacttcagc tgcctcatcc
     1681 cactcaatgt gctttttgta gaagatggca aaatggagcg ccaggtcttc cttgcaacat
     1741 ggaaggatat tcccaatgaa aatgaacttc agtttcagat taaggaatgt catttaaatg
     1801 ctgacactgt ttccagcaag ttgcaaaaca acaatgttta tactattgcc aagaggaatg
     1861 tggaagggca ggacatgctg taccaatccc tgaagctcac taatggcatt tggattttgg
     1921 ccgaactacg tatccagcca ggaaacccca attacacgct gtcactgaag tgtagagctc
     1981 ctgaagtctc tcaatacatc tatcaggtct acgacagcat tttgaaaaac ttaccggtca
     2041 tggtgtctaa gggcgaagag ctgatcaagg aaaatatgcg tatgaaggtg gtcatggaag
     2101 gttcggtcaa cggccaccaa ttcaaatgca caggtgaagg agaaggcaat ccgtacatgg
     2161 gaactcaaac catgaggatc aaagtcatcg agggaggacc cctgccattt gcctttgaca
     2221 ttcttgccac gtcgttcatg tatggcagcc gtacttttat caagtacccg aaaggcattc
     2281 ctgatttctt taaacagtcc tttcctgagg gttttacttg ggaaagagtt acgagatacg
     2341 aagatggtgg agtcgtcacc gtcatgcagg acaccagcct tgaggatggc tgtctcgttt
     2401 accacgtcca agtcagaggg gtaaactttc cctccaatgg tcccgtgatg cagaagaaga
     2461 ccaagggttg ggagcctaat acagagatga tgtatccagc agatggtggt ctgaggggat
     2521 acactcatat ggcactgaaa gttgatggtg gtggccatct gtcttgctct ttcgtaacaa
     2581 cttacaggtc aaaaaagacc gtcgggaaca tcaagatgcc cggtatccat gccgttgatc
     2641 accgcctgga aaggttagag gaaagtgaca atgaaatgtt cgtagtacaa cgcgaacacg
     2701 cagttgccaa gttcgccggg cttggtggtg ggatggacga gcgcggccgc gactctagat
     2761 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2821 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2881 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2941 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3001 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3061 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3121 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3181 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3241 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3301 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3361 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3421 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3481 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3541 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3601 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3661 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3721 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3781 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3841 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3901 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3961 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4021 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4081 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4141 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4201 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4261 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4321 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4381 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4441 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4501 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4561 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4621 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4681 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4741 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4801 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4861 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4921 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4981 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5041 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5101 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5161 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5221 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5281 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5521 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5821 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5881 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5941 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6001 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6061 tattaccgcc atgcat
//
