LOCUS Exported 9964 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION for baculovirus expression of CAMSAP2 in Sf9 cells. ACCESSION . VERSION . KEYWORDS pFasBac+GFP-CAMSAP2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9964) AUTHORS Hendershott MC, Vale RD TITLE Regulation of microtubule minus-end dynamics by CAMSAPs and Patronin. JOURNAL Proc Natl Acad Sci U S A. 2014 Apr 22;111(16):5860-5. doi: 10.1073/pnas.1404133111. Epub 2014 Mar 26. PUBMED 24706919 REFERENCE 2 (bases 1 to 9964) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9964 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 9..28 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 262..279 /label=L4440 /note="L4440 vector, forward primer" mobile_element 411..635 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (right end of the Tn7 transposon)" CDS complement(702..1235) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(1424..1452) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" promoter 1804..1895 /gene="polh from Autographa californica" /label=polyhedrin promoter /note="promoter for the baculovirus polyhedrin gene" primer_bind 1820..1837 /label=Polyhedrin-F /note="Polyhedrin promoter, forward primer" CDS 1962..1979 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2001..2021 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 2025..2738 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(2055..2083) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2678..2698 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" RBS 5785..5793 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" polyA_signal 7354..7488 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7391..7410) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7445..7464 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" mobile_element 7517..7682 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (left end of the Tn7 transposon)" rep_origin 7866..8321 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7953..7972) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8163..8184 /label=F1ori-F /note="F1 origin, forward primer" promoter 8348..8452 /gene="bla" /label=AmpR promoter CDS 8453..9313 /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(8671..8690) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9484..108 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 61 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 121 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 181 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 241 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg 301 cggtattttc tccttacgca tctgtgcggt atttcacacc gcagaccagc cgcgtaacct 361 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga 421 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag 481 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt 541 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga 601 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac 661 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg 721 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg 781 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca 841 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact 901 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc 961 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta 1021 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct 1081 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg 1141 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg 1201 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca 1261 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa 1321 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa 1381 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca 1441 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac 1501 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc 1561 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg 1621 cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc gcttgccggt 1681 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt 1741 gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc ggaatattaa 1801 tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta ttttactgtt 1861 ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca taccgtccca 1921 ccatcgggcg cggatctcgg tccgaaacca tgtcgtacta ccatcaccat caccatcacg 1981 attacgatat cccaacgacc gaaaacctgt attttcaggg cgccatgagc aaaggagaag 2041 aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca 2101 aattttctgt ccgtggagag ggtgaaggtg atgctacaaa cggaaaactc acccttaaat 2161 ttatttgcac tactggaaaa ctacctgttc cgtggccaac acttgtcact actctgacct 2221 atggtgttca atgcttttcc cgttatccgg atcacatgaa acggcatgac tttttcaaga 2281 gtgccatgcc cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggacct 2341 acaagacgcg tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa 2401 agggtattga ttttaaagaa gatggaaaca ttcttggaca caaactcgag tacaacttta 2461 actcacacaa tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca 2521 aaattcgcca caacgttgaa gatggttccg ttcaactagc agaccattat caacaaaata 2581 ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcg acacaatctg 2641 tcctttcgaa agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg 2701 ctgctgggat tacacatggc atggatgagc tctacaaagg atccggggat gctgcagacc 2761 ccagggagat gagaaagacg ttcattgttc cagccatcaa gccttttgac cactatgatt 2821 tctccagggc caaaatcgcc tgcaatctgg cctggctggt ggccaaagcc tttgggacag 2881 aaaatgtgcc agaggaactt caagaaccat tttacacaga tcagtatgac caggaacaca 2941 tcaaaccacc tgttgttaat ttgcttctat cggctgaact atactgtcgt gctgggagtc 3001 tcattctcaa gagtgatgct gcaaaacccc ttttgggcca tgatgctgta atccaggctt 3061 tagcacagaa aggtctttat gtcactgacc aggaaaaatt ggtaactgaa cgagatctcc 3121 acaagaaacc catacagatg agtgcacatt tggccatgat cgataccctc atgatggctt 3181 atactgtaga aatggtcagt atagaaaaag taattgcgtg tgctcagcag tattcagctt 3241 tttttcaagc cacagatctg ccctatgata ttgaggacgc tgtcatgtac tggataaata 3301 aggtaaatga acatttgaaa gacataatgg aacaagaaca aaaactgaaa gaacatcaca 3361 cagttgaagc tccaggaggt caaaagtctc cttccaaatg gttttggaaa ctggttccag 3421 ctcgttatcg gaaagagcaa acattgctta agcaactgcc ttgcattcca ttggtagaaa 3481 atttgttgaa ggatgggaca gatggctgtg cattagctgc ccttattcat ttttactgtc 3541 ctgatgttgt cagattagag gatatttgtt tgaaagaaac tatgtctttg gctgatagcc 3601 tgtataatct gcagctgatt caagaatttt gccaagaata cttgaaccag tgttgccatt 3661 tcactctgga agatatgctc tatgctgctt catccataaa gagtaattat ttggtgttca 3721 tggcggaact gttctggtgg tttgaagtgg tgaagccgtc ttttgtacag cctcgtgttg 3781 ttcgtccaca aggagctgaa cctgtaaaag atatgccttc aattcctgtc ttgaatgctg 3841 ccaaaagaaa tgtcttagat agtagttctg acttcccttc aagtggggaa ggagctacat 3901 ttacacagtc tcatcatcat ttgccttcta ggtattcacg tccccaggct cattcttcag 3961 cctcaggagg aattagaagg tcttcatcta tgtcttatgt tgatggcttc atagggacat 4021 ggcccaaaga gaaaagatca tcagtgcatg gcgtatcatt tgatatttct tttgataaag 4081 aagatagtgt acagagatcc actccaaacc gaggaatcac tcgttctatt agtaatgaag 4141 gacttactct gaacaacagt catgtatcta aacacattag gaaaaatttg tccttcaagc 4201 caataaatgg agaagaggaa gcagagagca ttgaagaaga acttaatata gattctcaca 4261 gtgacctcaa atcttgtgtg ccccttaaca caaatgaact aaattctaat gagaatattc 4321 attacaagct tccaaatgga gctttacaaa atagaatact tcttgacgag tttggcaatc 4381 agatcgagac accaagcatt gaagaagcat tacaaataat tcatgatact gaaaaatctc 4441 ctcatacacc tcagccagac caaattgcta atggcttctt tcttcatagt caagaaatga 4501 gtatcttaaa ttcaaatatc aagttaaatc aatctagtcc tgataatgta actgatacga 4561 aaggtgcctt gagtcccata actgacaata ctgaagtaga cactggaatt cacgttcctt 4621 cagaagatat tcctgaaact atggacgaag attcttcgtt gagagattat actgtaagct 4681 tggactctga catggatgat gcatctaaat ttcttcagga ttatgatatt cgaactggca 4741 acaccaggga agctttgagt ccttgtccaa gtactgtaag taccaagtct cagccaggca 4801 gcagtgcttc ttctagttct ggagttaaaa tgaccagctt tgctgaacaa aaattcagga 4861 aactgaatca taccgatgga aaaagtagtg gaagcagttc tcaaaaaact acaccagaag 4921 gctctgaact taatattcct catgtggttg cttgggcaca aattccagaa gaaacagggc 4981 ttccacaggg acgggacact acccagctgt tggcctctga aatggtgcat cttaggatga 5041 aactagaaga aaagaggcgt gctatagaag cccagaaaaa gaaaatggaa gctgctttta 5101 ccaaacagag acagaaaatg ggaaggacag cattccttac tgtagtgaaa aagaaagggg 5161 atgggatatc tcctctacga gaggaagcgg cgggtgcaga agatgagaaa gtatatactg 5221 atcgagcaaa agaaaaggaa tcacaaaaaa ctgatggaca aaggagcaag tcactggcag 5281 atataaaaga gagcatggag aatcctcaag ccaaatggct aaagtctcca actacaccta 5341 ttgatcctga gaagcagtgg aacctggcaa gcccctcaga agaaacttta aatgaaggag 5401 agattttaga atataccaaa tccattgaaa agttaaattc atccctgcat tttctacaac 5461 aagaaatgca acgcttgtca cttcagcagg agatgttaat gcagatgaga gagcaacaat 5521 cttgggtgat ttcacctcca caaccctctc cacagaaaca gattcgagat tttaagcctt 5581 ctaagcaggc aggcctgtca tcagccattg caccattctc ctcagactcc cctcgtccta 5641 ctcacccatc tccacagtct tctaacagga aaagtgcatc tttttctgtt aaaagtcaaa 5701 ggactcctag gccaaatgag ttaaaaataa cacctttgaa tcgaaccttg acacctcctc 5761 ggtctgtgga tagccttcct cggttaagga ggttttcacc aagtcaagtt cctattcaaa 5821 ctaggtcatt tgtatgtttt ggggatgatg gagaacctca gttaaaggaa tccaaaccta 5881 aagaggaagt taaaaaggag gaattggaat ccaaagggac tttggaacag cgtggacata 5941 atccagaaga aaaggaaatc aaaccttttg agtcaacagt ctctgaagtc ctatcactgc 6001 ctgtcacaga gactgtatgt ctgacaccaa atgaggacca attgaatcaa cccacagaac 6061 cccctcctaa acccgttttc ccacccactg ctccaaaaaa tgttaatctg attgaagttt 6121 ccctctcaga tttgaaaccc cctgaaaagg ctgatgtacc tgttgaaaaa tatgatggag 6181 aaagtgataa agaacaattt gatgatgacc agaaagtatg ctgtggattc ttttttaagg 6241 atgatcaaaa agcagaaaat gatatggcaa tgaaacgggc agctttgttg gagaaaagat 6301 taagaaggga aaaggaaact cagctccgga aacaacagtt ggaagcagaa atggagcata 6361 agaaggagga aacaaggcgt aaaactgagg aagaacgtca gaagaaagaa gatgagagag 6421 cacgcagaga atttattagg caagaatata tgaggcggaa acaactgaaa ctaatggaag 6481 atatggatac agtaattaaa ccccgtcctc aagtagtaaa acaaaaaaaa cagcgaccaa 6541 aatctattca cagagatcat attgaatccc ccaaaacacc aataaagggt cctccagtct 6601 ctagcctttc tttggcatcg ctgaacacgg gtgataacga gagtgtacat tcaggcaaga 6661 ggacgccaag atcagagtct gtagaaggct tcttatctcc aagtcgttgt ggcagtcgaa 6721 atggagaaaa agactgggag aatgcatcaa caacttcttc agtggcttct ggaacagaat 6781 atacaggacc aaagctctac aaagaaccca gtgcaaaatc caataagcac ataatacaaa 6841 atgctttagc tcattgctgt ttggctggaa aagtaaatga aggtcagaag aaaaaaatac 6901 tggaggaaat ggagaaatca gatgccaaca acttcttaat cttgttccgg gattcaggat 6961 gccagttcag atctttatac acttattgcc cagaaactga agaaatcaat aaactgactg 7021 ggataggccc taaatctatc actaaaaaaa tgattgaagg actttacaaa tataattctg 7081 acaggaaaca gtttagccac atacccgcta aaactttatc tgccagtgtt gatgcaatta 7141 ccattcatag ccatttatgg cagaccaaaa gaccagtaac acccaaaaaa cttttaccca 7201 ctaaggcata gggcatgcgg taccaagctt gtcgagaagt actagaggat cataatcagc 7261 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac 7321 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt 7381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 7441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt 7501 gagcctagga gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt 7561 aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct 7621 aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca 7681 cagcggggca tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga 7741 caaaccgtca tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc 7801 ttcgttatta atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga 7861 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 7921 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 7981 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 8041 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 8101 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 8161 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 8221 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 8281 atttaacgcg aattttaaca aaatattaac gtttacaatt tcaggtggca cttttcgggg 8341 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct 8401 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat 8461 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 8521 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg 8581 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 8641 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 8701 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 8761 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 8821 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc 8881 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 8941 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 9001 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca 9061 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 9121 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 9181 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 9241 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 9301 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact 9361 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 9421 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 9481 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 9541 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 9601 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 9661 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 9721 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 9781 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 9841 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga 9901 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 9961 ggag //