LOCUS Exported 8871 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bicaudal D protein aa1-594 (excluding start Met) tagged with N-terminal eGFP and flag and a C-terminal FKBP. ACCESSION . VERSION . KEYWORDS pBa-eGFP-flag-BicD2 594-FKBP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8871) 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 8871) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8871 /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..2012 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELY" CDS 1299..2012 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELY" primer_bind complement(1344..1365) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1605..1624) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1952..1973 /label=EGFP-C /note="EGFP, forward primer" CDS 2025..2048 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 3852..4172 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" polyA_signal 4292..4425 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 4382..4401 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4457..4473) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4457..4473) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4470..4492) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4481..4497 /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(4505..4535) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4550..4571 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4688..4705) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4859..5447) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4939..4958) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5618..6478) /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 6241..6260 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6479..6583) /gene="bla" /label=AmpR promoter primer_bind 6651..6669 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6707..6729) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6829..6848 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7042..7064 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7056..7073 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7057..7073 /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 ggcgaggagc 1321 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt 1381 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 1441 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg 1501 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg 1561 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca 1621 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg 1681 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca 1741 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga 1801 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc 1861 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc 1921 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg 1981 ccgggatcac tctcggcatg gacgagctgt acatggatat catggattac aaggatgacg 2041 atgacaaatc tcattccgcg ccgtcggagg aggaggagta tgcgcgtctg gtgatggagg 2101 cgcagccgga gtggctgcgc gccgaggtga agcggctgtc ccacgagctg gccgagacca 2161 cgcgtgagaa gatccaggcg gccgagtatg gactggcggt gctggaggag aagcaccagc 2221 tcaagctgca gttcgaggag ctcgaggtgg actatgaggc catccgcagc gagatggagc 2281 agctcaaaga ggcatttggc caggcacata caaaccacaa gaaggtggct gctgatggtg 2341 agagccggga ggagagcctg atccaggagt cggcctccaa ggagcagtac tacgtgcgga 2401 aggtgctgga gctgcagaca gagctgaaac agctgcgcaa cgtcctcacc aacacccagt 2461 ctgagaatga gcgcctcacg tctgtggccc aggagctaaa agagatcaac cagaatgtgg 2521 agatccagcg tggtcgcctg cgggatgaca tcaaggagta caagttccgg gaggcccgtc 2581 tacttcagga ctactcagag ctggaggagg agaacatcag cctgcagaaa caagtgtctg 2641 tgctcaggca gaaccaggtg gagtttgagg gcctcaagca tgaaatcaag cgcctggagg 2701 aggagacaga gtacctcaac agccagctag aggatgccat ccggcttaag gagatctctg 2761 aacggcagtt ggaggaggcg ctggagacac tgaagacaga gcgggagcag aagaacaacc 2821 tgcgcaagga gttgtcgcac tacatgagca tcaacgattc cttctatacc agccacctgc 2881 aggtctcctt agatggcctc aagttcagtg atgatactgt caccgcagag cccaacaatg 2941 acgccgaagc cctggtcaat ggctttgagc acagtggctt ggtcaaatca tccttggaca 3001 acaagacatc cacacccagg aaggatggcc tagctccacc ctcccccagc ctcgtctctg 3061 acctgctcag tgagctccac atatctgaga tccagaagct gaaacagcag ctggtgcaga 3121 tggagcggga gaaggtgggc ctgctggcaa cactgcagga cacacagaag cagctggagc 3181 aggctcgggg aaccctctct gagcagcatg agaaggtgaa tcgtctcaca gagaacctta 3241 gtgccctccg gcgcctgcag gctggcaagg agcgacagac ctcgttggat aatgagaagg 3301 accgtgacag ccacgaagac ggtgactact atgaggtgga catcaatggg cctgagatcc 3361 tggcctgcaa gtaccacgtg gctgtggctg aggctggcga gctccgggag cagctcaagg 3421 cgttgcgcag cacacacgaa gctcgcgaag cccagcacgc agaagaaaag ggccggtatg 3481 aggctgaggg ccaggcactc actgagaaga tctctctgct ggagaaggct agccaccagg 3541 accgagagct gctggcccat ctggaaaagg agctgaagaa ggtgagcgat gtggctggtg 3601 agacccaggg cagcctgaat gtggctcagg atgagctggt gaccttcagc gaggagctgg 3661 ccaacctcta ccaccatgtg tgcatgtgca acaacgagac gcccaaccgt gtcatgctcg 3721 actattatcg tgagggccag ggcaaggctg gccgcaccag cccagagggc cgcgggcgcc 3781 ggtcacctgt cctcttgccc aaggggctgt tggccacaga ggtgggccga gcagatcgga 3841 ccgtctctag aggagtgcag gtggaaacca tctccccagg agacgggcgc accttcccca 3901 agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaatttg 3961 attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc 4021 gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat 4081 ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca catgccactc 4141 tcgtcttcga tgtggagctt ctaaaactgg aaactagtta ttgaggatca taatcagcca 4201 tatcacatct gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct 4261 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta 4321 caaataaagc aatagcatca caaattcaca aataaagcat ttttttcact gcattctagt 4381 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccccgg gtaccgagct 4441 cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 4501 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 4561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 4681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 4801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 5041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 5101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 5161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 5221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 5281 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 5341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 5401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 5461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 5521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 5581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 5641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 5701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 5761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 6001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 6061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 6121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 6181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 6241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 6301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 6361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 6421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 6481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 6541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 6601 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 6661 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 6721 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 6781 agggcgcgtc agcgggtgtt ggcgggtgtc ggggttggct taactatgcg gcatcagagc 6841 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 6901 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 6961 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 7021 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagt 7081 tgggatcttt gcattggccc acggctctca ggatggggat gctccccttc agcacccggt 7141 tccccttgga aactgatggt cctggctctg tggcatggca gtggcactgt gaggagcccc 7201 taccagcagc acacagtggg tttggcactg ccacgctccg gatgccgcgc tctgatccaa 7261 ccccataatc aagggaaccc gaattgcccc atcattgccc ccaccacccc catcctgccg 7321 ggccctcaca ccccacgctg ccttgtggtg acattcccca gcccaaaccc acggcttcat 7381 ggctaccgcg gggcatttcc cattgccgcc ccattatcag ctctgcacac ctcccgctgt 7441 acccatgcct cgtggctgcc cttctttgac gtataatctt ctaattaata cccggccttg 7501 tcaaagtgga gcacaaacgt taattaattc cccagcaggc aggtaattaa cagtgtgact 7561 ccctttttgc tgcgagtggg gctgatacag agagatgtgg cactatggag cccacggggt 7621 cctggcactg ggtgcccacg gaggtcccca tgtgctgcag tgtcaccgcc tccgaggtga 7681 cagtattgtc cctgcggtgt ccctgcagct cagctctgtc cacagggcca cctccagttt 7741 ggaggggaca caatgcagcc ccgatgcaac ccatcctcgc agcatcccag ggacaaagac 7801 cccactgcaa gaccgcacac agggctgggt cccgctcccc taatatctac agtgcttttg 7861 catggcccct taatcaatgc agttaatcag catgcgctca tgcaccgctc tggagctgca 7921 aagcccctcg cagcgctgct caccaacacc gcgcaccgcc ccggcccagc ctgcagcacg 7981 cgctgcaaac aggaaagaaa caaaatattg cccaaatgta ggcaaaggca ttcggctgcc 8041 ttgacctccg ccgggccggg ccctgcctga ctcagctcct tactcagcgc tcgcttcctc 8101 cctccggctg ccaccgccgc agcgcacacc ctgacaaaga gtggccctta acgggctctg 8161 aggtgcaccc agcagtgcac tcagcagtcc aagggccggc ctggaggttt gcaccgctac 8221 gtgctgacat tagcattgaa cttggccctg ggtagtgctg caggccgggc ggggtgggtg 8281 tagagagtgc agcgcgcgtt gcacccggtg ccccttcccc tcccttgcat cccagcaggc 8341 tgcaccccag caccaggccc gtgcatgcat gctcctggtg ttattgcagc ctggtgcatg 8401 catgcgtctt agtggtgcag cgctgtgcat gcatcctcct tggtgtgtag cagcttagtg 8461 catgcatacc cctcggtgtt attgctgctc tgtgcacgca cgctcattgt atcacttcat 8521 cccagtgcat gcactcacac tggagcgatt gctgctcggt gcacgcacac tcattgtatc 8581 acgtcagctc agtggctgca cgcacaccgg tgttattgct gctcggtgcg tgcatgcaca 8641 tcagtgtcgc tgcagctcag tgcatgcacg ctcattgccc atcgctatcc ctgcctctcc 8701 tgctggcgct ccccgggagg tgacttcaag gggaccgcag gaccacctcg ggggtggggg 8761 gagggctgca cacgcggacc ccgctccccc tccccaacaa agcactgtgg aatcaaaaag 8821 gggggagggg ggatggaggg gcgcgtcaca cccccgcccc acaccctcac c //