LOCUS       sequence-376107-        7564 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DCX fusion to V5-tagged TurboID on 
            microtubules (DCX as the microtubule-targeting tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7564)
  AUTHORS   Cao X, Shami Shah A, Sanford EJ, Smolka MB, Baskin JM
  TITLE     Proximity Labeling Reveals Spatial Regulation of the 
            Anaphase-Promoting Complex/Cyclosome by a Microtubule Adaptor.
  JOURNAL   ACS Chem Biol. 2022 Aug 11. doi: 10.1021/acschembio.2c00527.
  PUBMED    35952650
REFERENCE   2  (bases 1 to 7564)
  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: "ACS Chem 
            Biol. 2022 Aug 11. doi: 10.1021/acschembio.2c00527."
COMMENT     Sequence Label: pCDNA3-DCX-TurboID-V5
FEATURES             Location/Qualifiers
     source          1..7564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        115..494
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        495..698
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     649..669
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     743..762
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        743..761
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      775..784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1945..2901
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             2908..2949
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        complement(2997..3015)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2998..3015)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3035..3052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3041..3265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3311..3739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3398..3417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3608..3629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3748..3768)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3753..4082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3933..4068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3995..4014
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4149..4943
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4203..4222)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4813..4832
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5117..5238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5154..5173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5208..5227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5287..5303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5287..5303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5300..5322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5311..5327
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5335..5341,5342..5359,5360..5365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5380..5401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5518..5535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5689..6277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5769..5788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6448..7239,7240..7308))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7309..7413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7488..7507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
       61 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      121 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      181 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      241 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      301 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      361 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      421 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      481 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      541 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      601 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      661 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      721 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
      781 atggaacttg attttggaca ctttgacgaa agagataaga catccaggaa catgcgaggc
      841 tcccggatga atgggttgcc tagccccact cacagcgccc actgtagctt ctaccgaacc
      901 agaaccttgc aggcactgag taatgagaag aaagccaaga aggtacgttt ctaccgcaat
      961 ggggaccgct acttcaaggg gattgtgtac gctgtgtcct ctgaccgttt tcgcagcttt
     1021 gacgccttgc tggctgacct gacgcgatct ctgtccgaca acatcaacct gcctcaggga
     1081 gtgcgttaca tttacaccat tgatggatcc aggaagatcg gaagcatgga tgaactggag
     1141 gaaggggaaa gctatgtctg ttcctcagac aacttcttta aaaaggtgga gtacaccaag
     1201 aatgtcaatc ccaactggtc tgtcaacgta aaaacatctg ccaatatgaa agccccccag
     1261 tctttggcta gcagcaacag tgcacaggcc agggagaaca aggactttgt gcgccccaag
     1321 ctggttacca tcatccgcag tggggtgaag cctcggaagg ctgtgcgtgt gcttctgaac
     1381 aagaagacag cccactcttt tgagcaagtc ctcactgata tcacagaagc catcaaactg
     1441 gagaccgggg ttgtcaaaaa actctacact ctggatggaa aacaggtaac ttgtctccat
     1501 gatttctttg gtgatgatga tgtgtttatt gcctgtggtc ctgaaaaatt tcgctatgct
     1561 caggatgatt tttctctgga tgaaaatgaa tgccgagtca tgaagggaaa cccatcagcc
     1621 acagctggcc caaaggcatc cccaacacct cagaagactt cagccaagag ccctggtcct
     1681 atgcgccgaa gcaagtctcc agctgactca ggtaacgacc aagacgcaaa cggaacctcc
     1741 agcagccagc tctctacccc caagtctaag cagtctccca tctctacgcc caccagtcct
     1801 ggcagcctcc ggaagcacaa ggacctgtac ctgcctctgt ccttggatga ctcggactcg
     1861 cttggtgatt ccatgagcgg cggccgcgga tcaggaagcg gatcaggaag cggatcagga
     1921 agcggatcag gaagcggatc aggaaaagac aatactgtgc ctctgaagct gatcgctctc
     1981 ctggctaatg gcgagttcca tagtggcgaa cagctgggag aaaccctggg catgtccagg
     2041 gccgctatca acaagcacat tcagactctg cgcgactggg gcgtggacgt gttcaccgtg
     2101 cccggaaagg gctactctct gcccgagcct atcccgctgc tgaacgctaa acagattctg
     2161 ggacagctgg acggcgggag cgtggcagtc ctgcctgtgg tcgactccac caatcagtac
     2221 ctgctggatc gaatcggcga gctgaagagt ggggatgctt gcattgcaga atatcagcag
     2281 gcagggagag gaagcagagg gaggaaatgg ttctctcctt ttggagctaa cctgtacctg
     2341 agtatgtttt ggcgcctgaa gcggggacca gcagcaatcg gcctgggccc ggtcatcgga
     2401 attgtcatgg cagaagcgct gcgaaagctg ggagcagaca aggtgcgagt caaatggccc
     2461 aatgacctgt atctgcagga tagaaagctg gcaggcatcc tggtggagct ggccggaata
     2521 acaggcgatg ctgcacagat cgtcattggc gccgggatta acgtggctat gaggcgcgtg
     2581 gaggaaagcg tggtcaatca gggctggatc acactgcagg aagcagggat taacctggac
     2641 aggaatactc tggccgctac gctgatccga gagctgcggg cagccctgga actgttcgag
     2701 caggaaggcc tggctccata tctgccacgg tgggagaagc tggataactt catcaataga
     2761 cccgtgaagc tgatcattgg ggacaaagag attttcggga ttagccgggg gattgataaa
     2821 cagggagccc tgctgctgga acaggacgga gttatcaaac cctggatggg cggagaaatc
     2881 agtctgcggt ctgccgaaaa ggctagcggc aagcccatcc ccaaccccct gctgggcctg
     2941 gacagcacct aatgaggatc cggcaagccc atccccaacc ccctgctggg ccctattcta
     3001 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3061 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3121 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3181 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3241 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3301 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3361 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3421 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3481 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3541 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3601 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3661 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3721 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4021 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4081 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4141 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4201 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4261 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4321 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4381 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4441 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4501 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4561 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4621 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4681 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4741 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4801 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4861 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4921 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4981 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5041 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5101 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5161 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5221 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5281 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5521 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5581 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5641 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5701 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5761 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5821 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5881 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5941 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6001 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6061 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6121 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7321 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7381 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7441 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     7501 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7561 tgcg
//
