LOCUS Exported 9582 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bicaudal D protein aa1-594 (excluding start Met) with N-terminal tandem dimer Tomato and flag tags and C-terminal FKBP. ACCESSION . VERSION . KEYWORDS pBa-tdTomato-flag-BicD2 594-FKBP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9582) AUTHORS Bentley M, Decker H, Luisi J, Banker G TITLE A novel assay reveals preferential binding between Rabs, kinesins, and specific endosomal subpopulations. JOURNAL J Cell Biol. 2015 Feb 2;208(3):273-281. Epub 2015 Jan 26. PUBMED 25624392 REFERENCE 2 (bases 1 to 9582) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9582 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..276 /label=chicken beta-actin promoter primer_bind 1209..1231 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" CDS 1299..2723 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELY" CDS 2736..2759 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 4563..4883 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" polyA_signal 5003..5136 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 5093..5112 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5168..5184) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5168..5184) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5181..5203) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5192..5208 /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(5216..5246) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5261..5282 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5399..5416) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5570..6158) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5650..5669) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6329..7189) /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 6952..6971 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7190..7294) /gene="bla" /label=AmpR promoter primer_bind 7362..7380 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7418..7440) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7540..7559 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7753..7775 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7767..7784 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7768..7784 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa 61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg 121 cgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg 181 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg 241 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgtt gccttcgccc 301 cgtgccccgc tccgcgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc 361 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa 421 tgacggctcg tttcttttct gtggctgcgt gaaagcctta aagggctccg ggagggccct 481 ttgtgcgggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc 541 gtgcggcccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg 601 ctccgcgtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc gggggggctg 661 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg 721 cgcggcggtc gggctgtaac ccccccctgc acccccctcc ccgagttgct gagcacggcc 781 cggcttcggg tgcggggctc cgtgcggggc gtggcgcggg gctcgccgtg ccgggcgggg 841 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg 901 gggaggggcg cggcggcccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca 961 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctggcg 1021 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcgggcg aagcggtgcg 1081 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc 1141 ttctccatct ccagcctcgg ggctgccgca gggggacggc tgccttcggg ggggacgggg 1201 cagggcgggg ttcggcttct ggcgtgtgac cggcggggtt tatatcttcc cttctctgtt 1261 cctccgcagc ccccaagctt ggcgcgccta ggcctaccat ggtgagcaag ggcgaggagg 1321 tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt 1381 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga 1441 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt 1501 acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct 1561 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg 1621 tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca 1681 ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca 1741 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc 1801 tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg 1861 tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg 1921 actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctggggc 1981 atggcaccgg cagcaccggc agcggcagct ccggcaccgc ctcctccgag gacaacaaca 2041 tggccgtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc 2101 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca 2161 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt 2221 tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc 2281 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg 2341 tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc 2401 gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg 2461 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc 2521 tgaagctgaa ggacggcggc cgctacctgg tggagttcaa gaccatctac atggccaaga 2581 agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca 2641 acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc 2701 tgtacggcat ggacgagctg tacatggata tcatggatta caaggatgac gatgacaaat 2761 ctcattccgc gccgtcggag gaggaggagt atgcgcgtct ggtgatggag gcgcagccgg 2821 agtggctgcg cgccgaggtg aagcggctgt cccacgagct ggccgagacc acgcgtgaga 2881 agatccaggc ggccgagtat ggactggcgg tgctggagga gaagcaccag ctcaagctgc 2941 agttcgagga gctcgaggtg gactatgagg ccatccgcag cgagatggag cagctcaaag 3001 aggcatttgg ccaggcacat acaaaccaca agaaggtggc tgctgatggt gagagccggg 3061 aggagagcct gatccaggag tcggcctcca aggagcagta ctacgtgcgg aaggtgctgg 3121 agctgcagac agagctgaaa cagctgcgca acgtcctcac caacacccag tctgagaatg 3181 agcgcctcac gtctgtggcc caggagctaa aagagatcaa ccagaatgtg gagatccagc 3241 gtggtcgcct gcgggatgac atcaaggagt acaagttccg ggaggcccgt ctacttcagg 3301 actactcaga gctggaggag gagaacatca gcctgcagaa acaagtgtct gtgctcaggc 3361 agaaccaggt ggagtttgag ggcctcaagc atgaaatcaa gcgcctggag gaggagacag 3421 agtacctcaa cagccagcta gaggatgcca tccggcttaa ggagatctct gaacggcagt 3481 tggaggaggc gctggagaca ctgaagacag agcgggagca gaagaacaac ctgcgcaagg 3541 agttgtcgca ctacatgagc atcaacgatt ccttctatac cagccacctg caggtctcct 3601 tagatggcct caagttcagt gatgatactg tcaccgcaga gcccaacaat gacgccgaag 3661 ccctggtcaa tggctttgag cacagtggct tggtcaaatc atccttggac aacaagacat 3721 ccacacccag gaaggatggc ctagctccac cctcccccag cctcgtctct gacctgctca 3781 gtgagctcca catatctgag atccagaagc tgaaacagca gctggtgcag atggagcggg 3841 agaaggtggg cctgctggca acactgcagg acacacagaa gcagctggag caggctcggg 3901 gaaccctctc tgagcagcat gagaaggtga atcgtctcac agagaacctt agtgccctcc 3961 ggcgcctgca ggctggcaag gagcgacaga cctcgttgga taatgagaag gaccgtgaca 4021 gccacgaaga cggtgactac tatgaggtgg acatcaatgg gcctgagatc ctggcctgca 4081 agtaccacgt ggctgtggct gaggctggcg agctccggga gcagctcaag gcgttgcgca 4141 gcacacacga agctcgcgaa gcccagcacg cagaagaaaa gggccggtat gaggctgagg 4201 gccaggcact cactgagaag atctctctgc tggagaaggc tagccaccag gaccgagagc 4261 tgctggccca tctggaaaag gagctgaaga aggtgagcga tgtggctggt gagacccagg 4321 gcagcctgaa tgtggctcag gatgagctgg tgaccttcag cgaggagctg gccaacctct 4381 accaccatgt gtgcatgtgc aacaacgaga cgcccaaccg tgtcatgctc gactattatc 4441 gtgagggcca gggcaaggct ggccgcacca gcccagaggg ccgcgggcgc cggtcacctg 4501 tcctcttgcc caaggggctg ttggccacag aggtgggccg agcagatcgg accgtctcta 4561 gaggagtgca ggtggaaacc atctccccag gagacgggcg caccttcccc aagcgcggcc 4621 agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaattt gattcctccc 4681 gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc cgaggctggg 4741 aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata tctccagatt 4801 atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact ctcgtcttcg 4861 atgtggagct tctaaaactg gaaactagtt attgaggatc ataatcagcc atatcacatc 4921 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa 4981 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag 5041 caatagcatc acaaattcac aaataaagca tttttttcac tgcattctag ttgtggtttg 5101 tccaaactca tcaatgtatc ttatcatgtc tggatccccg ggtaccgagc tcgaattcgt 5161 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5221 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5281 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5341 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 5401 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 5461 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 5521 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 5581 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 5641 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 5701 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 5761 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 5821 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 5881 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 5941 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6001 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6061 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 6121 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6181 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6241 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6301 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6361 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6421 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6481 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 6541 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 6601 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6661 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6721 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6781 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6841 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6901 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 6961 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 7021 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7081 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7141 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7201 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7261 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7321 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 7381 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 7441 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 7501 cagcgggtgt tggcgggtgt cggggttggc ttaactatgc ggcatcagag cagattgtac 7561 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca 7621 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct 7681 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa 7741 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ttgggatctt 7801 tgcattggcc cacggctctc aggatgggga tgctcccctt cagcacccgg ttccccttgg 7861 aaactgatgg tcctggctct gtggcatggc agtggcactg tgaggagccc ctaccagcag 7921 cacacagtgg gtttggcact gccacgctcc ggatgccgcg ctctgatcca accccataat 7981 caagggaacc cgaattgccc catcattgcc cccaccaccc ccatcctgcc gggccctcac 8041 accccacgct gccttgtggt gacattcccc agcccaaacc cacggcttca tggctaccgc 8101 ggggcatttc ccattgccgc cccattatca gctctgcaca cctcccgctg tacccatgcc 8161 tcgtggctgc ccttctttga cgtataatct tctaattaat acccggcctt gtcaaagtgg 8221 agcacaaacg ttaattaatt ccccagcagg caggtaatta acagtgtgac tccctttttg 8281 ctgcgagtgg ggctgataca gagagatgtg gcactatgga gcccacgggg tcctggcact 8341 gggtgcccac ggaggtcccc atgtgctgca gtgtcaccgc ctccgaggtg acagtattgt 8401 ccctgcggtg tccctgcagc tcagctctgt ccacagggcc acctccagtt tggaggggac 8461 acaatgcagc cccgatgcaa cccatcctcg cagcatccca gggacaaaga ccccactgca 8521 agaccgcaca cagggctggg tcccgctccc ctaatatcta cagtgctttt gcatggcccc 8581 ttaatcaatg cagttaatca gcatgcgctc atgcaccgct ctggagctgc aaagcccctc 8641 gcagcgctgc tcaccaacac cgcgcaccgc cccggcccag cctgcagcac gcgctgcaaa 8701 caggaaagaa acaaaatatt gcccaaatgt aggcaaaggc attcggctgc cttgacctcc 8761 gccgggccgg gccctgcctg actcagctcc ttactcagcg ctcgcttcct ccctccggct 8821 gccaccgccg cagcgcacac cctgacaaag agtggccctt aacgggctct gaggtgcacc 8881 cagcagtgca ctcagcagtc caagggccgg cctggaggtt tgcaccgcta cgtgctgaca 8941 ttagcattga acttggccct gggtagtgct gcaggccggg cggggtgggt gtagagagtg 9001 cagcgcgcgt tgcacccggt gccccttccc ctcccttgca tcccagcagg ctgcacccca 9061 gcaccaggcc cgtgcatgca tgctcctggt gttattgcag cctggtgcat gcatgcgtct 9121 tagtggtgca gcgctgtgca tgcatcctcc ttggtgtgta gcagcttagt gcatgcatac 9181 ccctcggtgt tattgctgct ctgtgcacgc acgctcattg tatcacttca tcccagtgca 9241 tgcactcaca ctggagcgat tgctgctcgg tgcacgcaca ctcattgtat cacgtcagct 9301 cagtggctgc acgcacaccg gtgttattgc tgctcggtgc gtgcatgcac atcagtgtcg 9361 ctgcagctca gtgcatgcac gctcattgcc catcgctatc cctgcctctc ctgctggcgc 9421 tccccgggag gtgacttcaa ggggaccgca ggaccacctc gggggtgggg ggagggctgc 9481 acacgcggac cccgctcccc ctccccaaca aagcactgtg gaatcaaaaa ggggggaggg 9541 gggatggagg ggcgcgtcac acccccgccc cacaccctca cc //