LOCUS       sequence-342225-        6177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Dok1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6177)
  AUTHORS   Yu CH, Rafiq NB, Cao F, Zhou Y, Krishnasamy A, Biswas KH, Ravasio A,
            Chen Z, Wang YH, Kawauchi K, Jones GE, Sheetz MP
  TITLE     Integrin-beta3 clusters recruit clathrin-mediated endocytic 
            machinery in the absence of traction force.
  JOURNAL   Nat Commun. 2015 Oct 28;6:8672. doi: 10.1038/ncomms9672.
  PUBMED    26507506
REFERENCE   2  (bases 1 to 6177)
  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: "2015-10-28"; volume: "6:8672. doi"; pages: " 
            10.1038/ncomms9672"
COMMENT     Sequence Label: Dok1-EGFP
FEATURES             Location/Qualifiers
     source          1..6177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    621..645
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2092..2116)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2136..2145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2142..2144,2145..2147,2148..2861)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2142..2144,2145..2147,2148..2858)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2187..2208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2448..2467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2795..2816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2984..3105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3021..3040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3075..3094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3112..3567)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3249..3270)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3461..3480
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3594..3698
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3700..4057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3700..3891
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3723..3743)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3908..4043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4092..4886
                     /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             4092..4886
                     /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(4146..4165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4756..4775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5074..5093)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5118..5165
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5494..6082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5983..6002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      601 gtcagatccg ctagcatcaa acaagtttgt acaaaaaagt tggcaccatg gacggagcag
      661 tgatggaagg gccgcttttt ttgcagagtc agcgctttgg gaccaagagg tggaggaaga
      721 cctgggccgt gctctacccg gccagtcccc acggcgtagc gcggctcgag ttctttgacc
      781 ataaggggtc gagctctggg ggtggccgag ggagctcgcg ccgcctggac tgcaaagtga
      841 tccgtctggc tgagtgtgtg agtgtggccc ccgtcaccgt ggagaccccc cctgagcccg
      901 gcgccactgc cttccgcctg gacactgctc agcgctcgca cctgctggcg gccgacgcgc
      961 cgtccagtgc agcctgggtg cagacgctgt gccgaaacgc ctttccgaaa ggcagctgga
     1021 ctctggcgcc taccgataac ccacctaagc tttctgccct ggagatgctg gagaactcct
     1081 tgtacagccc tacctgggaa ggatcccaat tctgggtaac ggtgcagagg actgaggccg
     1141 ccgagcgctg tggcctgcat ggctcctacg tgctgagggt ggaggctgaa aggctgactc
     1201 tcctgaccgt gggggcccag agtcagatac tggagccact cctgtcctgg ccctacactc
     1261 tgttgcgtcg ctatggccgg gacaaggtca tgttctcttt cgaggccggc cgccgctgcc
     1321 cctcaggccc tggaaccttc accttccaga cggcacaggg aaatgacatc ttccaggcag
     1381 ttgagactgc catccaccgg cagaaggccc agggaaaggc cggacagggg cacgatgttc
     1441 tcagagctga ctcccatgaa ggggaggtgg cagaggggaa gttgccttcc ccacctggcc
     1501 cccaagagct cctcgacagt cccccagccc tgtatgctga gcccttagac tccctgcgca
     1561 ttgctccatg cccttcccag gactccctat actcagaccc cttggacagc acgtctgctc
     1621 aggcaggaga gggagtacaa cggaagaaac ctctctattg ggacttgtat gagcatgcgc
     1681 agcagcagtt gctgaaggcc aagctgacag accccaaaga ggatcccatc tatgatgaac
     1741 ctgagggcct ggccccagtc cctccccagg gcctttatga tctgcctcgg gagcccaagg
     1801 atgcatggtg gtgccaagct cgggtgaagg aggagggcta tgagctcccc tacaaccctg
     1861 ccactgatga ctacgctgtg ccaccccctc ggagcacaaa gcccctcctt gctcccaagc
     1921 cccagggccc agccttccct gaacctggta ctgcaactgg cagtggcatc aaaagccaca
     1981 actcagccct gtacagccag gtccagaaga gcggggcctc agggagctgg gactgtgggc
     2041 tctctagagt agggactgac aagactgggg tcaagtcaga gggctctacc ttgccaactt
     2101 tcttgtacaa agtggttcga tgggatccac cggtcgccac catggtgagc aagggcgagg
     2161 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2221 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2281 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2341 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2401 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2461 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2521 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2581 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2641 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2701 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2761 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2821 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat
     2881 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2941 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3001 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3061 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3121 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3181 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3241 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3301 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3361 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3421 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3481 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3541 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3601 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3661 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3721 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3781 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3841 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3901 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3961 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4021 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4081 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4141 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4201 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4261 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4321 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4381 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4441 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4501 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4561 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4621 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4681 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4741 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4801 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4861 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4921 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4981 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5041 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5101 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5161 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5221 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5281 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5341 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5401 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5641 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataa
//
