LOCUS       sequence-463039-        7622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human dynein subunit DIC1 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7622)
  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 7622)
  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: pcDNA FRT to BioID-cDIC1-3xFlag
FEATURES             Location/Qualifiers
     source          1..7622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     475..494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     728..745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    862..883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(898..903,904..921,922..928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    936..952
                     /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     941..963
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     960..976
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     960..976
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1025..1146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1036..1055)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1090..1109
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1276..2298)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(1276..2298)
                     /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(2306..2353)
                     /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     2485..2504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2637..2861
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2868..2933)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(2934..3632)
                     /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(5559..5578)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5560..5578)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5623..5826)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5652..5672)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5827..6206
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6378..6397
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6472..6576
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6577..6645,6646..7437)
                     /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(6795..6814)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7608..7622,1..574)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
       61 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      121 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      361 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      601 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      661 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      721 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      781 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      841 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      901 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      961 aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg gtatacagac
     1021 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     1081 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1141 caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     1201 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     1261 atctcgtagt acgtgctatt cctttgccct cggacgagtg ctggggcgtc ggtttccact
     1321 atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc gggcgatttg
     1381 tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac cctgcgccca
     1441 agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg
     1501 gagcatatac gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta
     1561 gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt tggcgacctc
     1621 gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta tgcggccatt
     1681 gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga cttcggggca
     1741 gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac tgacggtgtc
     1801 gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata tgaaatcacg
     1861 ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct
     1921 aagatcggcc gcagcgatcg catccatggc ctccgcgacc ggctgcagaa cagcgggcag
     1981 ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga tgcaataggt
     2041 caggctctcg ctgaattccc caatgtcaag cacttccgga atcgggagcg cggccgatgc
     2101 aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat ttacccgcag
     2161 gacatatcca cgccctccta catcgaagct gaaagcacga gattcttcgc cctccgagag
     2221 ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc
     2281 ggtgagttca ggctttttgg ccaaggaagt tcctatactt tctagagaat aggaacttcg
     2341 gaataggaac ttctaggtac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2401 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2461 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2521 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2581 acagggcgcg tggggatacc ccctagagcc ccagctggtt ctttccgcct cagaagccat
     2641 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
     2701 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
     2761 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
     2821 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgttactt gtcatcgtca
     2881 tccttgtaat cgatgtcatg atctttataa tcaccgtcat ggtctttgta gtcgcttctt
     2941 ctcaggctga actcgccgct caggatctcc ttgatgccct cctcggtcag gatcagggcg
     3001 ccgcccttct cgctcaggcc caccagcttg ccggtgatct tgccctcgcc cagcagcttc
     3061 acctcctcgc ccaggtacag catcttgctc tcgatcttgc ccttgaactc cttgaagctc
     3121 ttctccttga acttcttcag gttctcgctg attctcttca gcaccttcag cagcacctcc
     3181 tttctgtccc agtccttgcc ggtgatctcg tacagggtgg tggctctgtc cttgatctcc
     3241 tcggggatct ctctctggtt cacgttgatg ccgatgccca cgatcagctt gtccttgctc
     3301 agctcgcaca gcacgccgct caccttcttc tcctggaagt acacgtcgtt gggccacttc
     3361 aggctgaagg ggatctcggt gatctcctcc agggcctcgc tcacgctcag gcccagcacc
     3421 aggggcagct gcagcaggtt ctcgaactcc ttggggttca gcaggaagct gaagtacagg
     3481 ccgccctcct ggctcagcca ctttctgccc aggccgcctc tgcccttggt ctgtctgtcg
     3541 gccaccaggg cggtgccgta gctcacgttc cactccttca gtctctcctg ggtgctgtcc
     3601 acctccttca gccagatcag gttcttgaac atggcaccgg ctccagcgcc tgcaccagct
     3661 ccggcagata actcaacagt gccttcctcc tcgctatcag ctctgttagc acgaatttcc
     3721 acaagggtcc tggcaaatcg ggtccattca tcattgtggg gaactgcaag ctctccaacg
     3781 tcatagaccc aaatacggcc ttccgagtcc ccaacagcaa cttctttgcc agcttgggcc
     3841 caacgaacac ggtttagggc ggatgccccc tcaatggcca cacttgctgt tggaacctcg
     3901 gtgtcattgt tgaggttcca gaggtccaag cgccccatcc cgtccacgca ggcaaaaagc
     3961 gcaggatgca cgggggacca catgacatcg tacacatagt ctgcattgtc ttcaaaggag
     4021 tagagcggct tgttgtgctt ggtggtccac agtttgacag tccagtcaaa tgatgatgtg
     4081 acaaacaggt gagaaaagtc gattggtccc actgccatgt ggcagttaat tcctgtcact
     4141 ggcccttggt gaccttcaaa gacctcacca atacctgctt tgcttccatg acgacaagcc
     4201 gtgtagactg taccttcctc actgccaacc acgaagttat tgacgtctcc cgttgggaaa
     4261 gccattccgg taacagcgac aggcttggac ttattgtaca ccagctccat gctctcctgt
     4321 ggagttgaga gcatgtccag gctccaggaa cacattttgc catcagtgga gacagtgatg
     4381 aggttatgag cattctgggt cccaacaaca tttacacagt acacgggatg cgtgtgtgca
     4441 gcagctgata agggtgtccg ctgcactgga gtccttcgat gactgcgatt gtcccagagg
     4501 acaatctggc ccgagtaagt cccaccaacc accaagttag gatggaaacg ggcgaagcag
     4561 accgacatca cagaggactg acagtggaag acgtattctg gtgtggtttt cttaaacttc
     4621 atgttccaaa ccaaggccac tccatctggt tcatggggag catcttcatt gttgttgtaa
     4681 gaagccacca tcagctcagg gtactggagg gaccagtcca tacaagtgac cactcgatgc
     4741 ttggaccaat gttcatcata gaactgacga ttgaaagaaa gattggctcc agcctgaaca
     4801 tccccatctt tttcctctaa ctctcggccg ctgtagtcaa aaaagatgtc ggaatcttca
     4861 gccagggctc tttcaattac ccgtattgtc cggtcaaaaa agatgagaaa ttcctctgaa
     4921 tgaaggatct gctgtttttc ttcctctgtc aactctcttg gaggggcttc cttcacttcc
     4981 tgttttttgt cctgattttc cagttctgag tcctggccaa ctttagattc caccatttcc
     5041 tcatcttcct catcctcttc agactgatgc gtggcaagag gagtctgggt ctcctttgag
     5101 taggacacta cttcccttgg caggaaatcc acttgggtga cctttgacac gcccagttta
     5161 tgcagtcttc ttccaagttc tgagtctgac tgcagctgga gcactgaggg gtctgtgtcc
     5221 cactgcaggg tccttgttaa tggccccaga tcgcctgagt cttggcttcc agcttcactg
     5281 ggagtgctca ctgatttcga ggagggagac ataggggttg ggactagagg cggctccggt
     5341 gagataccaa tgctttgcag caaagcctct gtctctcgtc gtttgcgatc cagatcagag
     5401 tcgtcctgaa cgggttcttt cttctgctgc atatcagcct ctttcttttt cctctcctct
     5461 tccttccgtt tcttctcttc tcttatctgt gctaagcgct gctttttgcg ctctagctca
     5521 gcttttaagt cacttttgtc agacatcagc ttgggtctcc ctatagtgag tcgtattaat
     5581 ttcgataagc cagtaagcag tgggttctct agttagccag agagctctgc ttatatagac
     5641 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     5701 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     5761 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     5821 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     5881 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     5941 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     6001 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     6061 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     6121 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     6181 ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat cgcgaagcag
     6241 cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc cttgccttgt
     6301 tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag cagggagcag
     6361 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataggggatc
     6421 gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6481 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6541 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6601 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6661 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6721 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6781 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6841 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6901 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6961 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7021 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7081 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7141 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7201 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7261 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7321 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7381 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7441 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7501 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7561 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7621 tt
//
