LOCUS       sequence-463038-        7574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human dynein subunit DIC2 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7574)
  AUTHORS   Redwine WB, DeSantis ME, Hollyer I, Htet ZM, Tran PT, Swanson SK, 
            Florens L, Washburn MP, Reck-Peterson SL
  TITLE     The human cytoplasmic dynein interactome reveals novel activators of
            motility.
  JOURNAL   Elife. 2017 Jul 18;6:e28257. doi: 10.7554/eLife.28257.
  PUBMED    28718761
REFERENCE   2  (bases 1 to 7574)
  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: "Elife."; 
            date: "2017-07-18"; volume: "6:e28257. doi"; pages: " 
            10.7554/eLife.28257"
COMMENT     Sequence Label: pcDNA5 FRT to BioID-nDIC2-3xFlag
FEATURES             Location/Qualifiers
     source          1..7574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1291..2313)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(1291..2313)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(2321..2368)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     2500..2519
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2652..2876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2883..2948)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      3475..3484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4815..5513)
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="MFKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKW
                     LSQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKV
                     SGVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISE
                     NLKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKE
                     ILSGEFSLRRS"
     primer_bind     complement(5526..5545)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5527..5545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5590..5793)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5619..5639)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5794..6173
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6345..6364
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6439..6543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6544..6612,6613..7404)
                     /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"
     primer_bind     complement(6762..6781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtagtacgtg ctattccttt gccctcggac gagtgctggg
     1321 gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggccgcgct
     1381 tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca
     1441 tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg
     1501 gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa gctccggatg
     1561 cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa
     1621 gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc
     1681 tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg
     1741 ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga
     1801 cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc
     1861 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa
     1921 cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg cgaccggctg
     1981 cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg
     2041 ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg
     2101 gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca
     2161 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc
     2221 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt
     2281 ctcgacagac gtcgcggtga gttcaggctt tttggccaag gaagttccta tactttctag
     2341 agaataggaa cttcggaata ggaacttcta ggtacgtgaa ccatcaccct aatcaagttt
     2401 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2461 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2521 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2581 gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc tggttctttc
     2641 cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
     2701 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     2761 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     2821 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcgt
     2881 tacttgtcat cgtcatcctt gtaatcgatg tcatgatctt tataatcacc gtcatggtct
     2941 ttgtagtcag caggtattcg ggtagctgct tcctcctctg catcagctcg gtttgcatta
     3001 atttctgcaa gtgttcggcc aaaccgtgcc cattcatcat tgcggggaac agcaatctgc
     3061 tctcccacat cgtatataac aatctgtcct tcagaatcac ccacagcaat ctctctgcca
     3121 gaatgggtcc atctcacacg attaagagca ggattaccct ccacagaaat gctggcagtt
     3181 ggtacctctg tgtcattatt gagattccac aaatccaatc tccccatgcc atccacacag
     3241 gcaaacaggg ctgggtgggt aggtgaccac ataacatcat aaacatagtc tgcattatct
     3301 tcaaatgaat acaaaggctt gttattctta gttgtccaaa gctttactgt ccagtcaaac
     3361 gatgaagtga caaaaagatg tgagaagtct actgctccaa cagctgcatg acaatggatg
     3421 ccagtgattg gtccttgatg cccctcaaac atctcactga ttccagcttt gctgccatgg
     3481 cggcatgctg tgtacacaga accttcttca ctcccaacaa caaagttgtt gacatctcca
     3541 acagggaagg acatagatgt cacagctact gcttttgact gtttatgaac caactccatg
     3601 ctatcctgtg gatgggaaag catgtccaga ctccatgaac aaatttttcc atcagtagag
     3661 atgctaatca gattgtgagc attttgtgtt ccaacaacat ttacacaata tacagggtgt
     3721 gtgtgtgcag ctgctgacag tggagttctt tgcactggag ttcttttatt gctacggtta
     3781 tcccaaagca caatttggcc tgaatatgta ccaccaacaa caagatttgg atgaaatttt
     3841 gcaaatgtgg cagacatcac agctgactgg cagtgaaaca catactctgg ggtagttttt
     3901 ttgtatttca tattccatac aagggccaca ccatcaggct catgaggggc atcttcattg
     3961 ttgttatagg aagccacgag taactccgga tactgagatg accaatccaa acaactaacc
     4021 acccgatgct ttgaccaacg ttcgtcaaaa aattgtcgat ttaatgacag tttagcacct
     4081 gcttgaatct ctccttcttt gtcttccaaa tctctcccac tatagtcaaa gaagatgtta
     4141 atctgctcag aaagagctct ttctacaatt cttgtagaat ggtcaaagaa acttaaaaat
     4201 tcctcagagt gcaagatttg ttgcttttct tcttcagtca gctcatgagg gggagcttta
     4261 ctatcatttt cctcatcttt ctttaaagtt ttctcttctt caggttcaat aggtggttta
     4321 ggagccacta catcatcatc ttcctcttca tcttctttgg gttgagccat aactggagtc
     4381 tgagtttcct ttgtatacgt gacaatttct cgaggaggaa agtcgacttg cgtgatttta
     4441 gccattccaa gtttaatagg tcctcgtcta gatcccacgg cgccatctcc agagtcttgg
     4501 cttccagctt cacttggagt gctcacagat ttggaggatg gagacatagg aggagggaca
     4561 atgggggatt ctggagttag ccccatgctt tgaagcaatg cttcagcttc tctccttttt
     4621 ttttcaagat ctgattcttc ttgcacagga gcaacagctt ccttcttctg gtctgtttct
     4681 ttttttttcc tttcttcttc ttttctcttc ttttcctctc tgatttgggc cagtcgctgc
     4741 ttcttacgtt ccaactcagc ctttaattca cttttgtctg acatggcacc ggctccagcg
     4801 cctgcaccag ctccgcttct tctcaggctg aactcgccgc tcaggatctc cttgatgccc
     4861 tcctcggtca ggatcagggc gccgcccttc tcgctcaggc ccaccagctt gccggtgatc
     4921 ttgccctcgc ccagcagctt cacctcctcg cccaggtaca gcatcttgct ctcgatcttg
     4981 cccttgaact ccttgaagct cttctccttg aacttcttca ggttctcgct gattctcttc
     5041 agcaccttca gcagcacctc ctttctgtcc cagtccttgc cggtgatctc gtacagggtg
     5101 gtggctctgt ccttgatctc ctcggggatc tctctctggt tcacgttgat gccgatgccc
     5161 acgatcagct tgtccttgct cagctcgcac agcacgccgc tcaccttctt ctcctggaag
     5221 tacacgtcgt tgggccactt caggctgaag gggatctcgg tgatctcctc cagggcctcg
     5281 ctcacgctca ggcccagcac caggggcagc tgcagcaggt tctcgaactc cttggggttc
     5341 agcaggaagc tgaagtacag gccgccctcc tggctcagcc actttctgcc caggccgcct
     5401 ctgcccttgg tctgtctgtc ggccaccagg gcggtgccgt agctcacgtt ccactccttc
     5461 agtctctcct gggtgctgtc cacctccttc agccagatca ggttcttgaa catcagcttg
     5521 ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg gttctctagt
     5581 tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     5641 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     5701 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     5761 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     5821 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     5881 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     5941 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     6001 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     6061 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     6121 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt
     6181 atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat
     6241 gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc
     6301 gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat
     6361 tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc
     6421 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6481 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6541 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6601 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6661 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6721 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6781 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6841 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6901 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6961 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7021 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7081 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7141 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7201 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7261 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7321 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7381 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7441 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7501 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7561 atcaaaggat cttc
//
