LOCUS Exported 5952 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses C-terminally HA-tagged Arf6(T27N) in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA3/hArf6(T27N)-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5952) AUTHORS Makyio H, Ohgi M, Takei T, Takahashi S, Takatsu H, Katoh Y, Hanai A, Ueda T, Kanaho Y, Xie Y, Shin HW, Kamikubo H, Kataoka M, Kawasaki M, Kato R, Wakatsuki S, Nakayama K TITLE Structural basis for Arf6-MKLP1 complex formation on the Flemming body responsible for cytokinesis. JOURNAL EMBO J. 2012 May 30;31(11):2590-603. doi: 10.1038/emboj.2012.89. Epub 2012 Apr 20. PUBMED 22522702 REFERENCE 2 (bases 1 to 5952) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5952 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 98..124 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(127..143) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(127..143) /label=pBluescriptSK /note="For pBluescript vector" promoter complement(153..171) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(154..171) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(191..208) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 197..421 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 467..895 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(554..573) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 764..785 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(904..924) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 909..1238 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1089..1224 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1151..1170 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 1305..2099 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1359..1378) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1969..1988 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 2273..2394 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2310..2329) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2364..2383 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(2443..2459) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2443..2459) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2456..2478) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2467..2483 /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(2491..2521) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2536..2557 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2674..2691) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2845..3433) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2925..2944) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3604..4464) /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 4227..4246 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4465..4569) /gene="bla" /label=AmpR promoter primer_bind complement(4644..4663) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 4835..5214 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5215..5418 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5369..5389 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 5463..5482 /label=T7 /note="T7 promoter, forward primer" promoter 5463..5481 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 5513..5522 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 ggacaggaac tggtatgtgc agccctcctg tgccacctca ggggacggac tctatgaggg 61 gctcacatgg ttaacctcta actacaaatc tctcgagtat ccatacgatg taccagacta 121 cgcagagatc cactagttct agagggccct attctatagt gtcacctaaa tgctagagct 181 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 241 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 301 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 361 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 421 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 481 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 541 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 601 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 661 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 721 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 781 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 841 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 901 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 961 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 1021 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 1081 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 1141 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 1201 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 1261 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 1321 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 1381 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 1441 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 1501 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 1561 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 1621 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 1681 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 1741 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 1801 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 1861 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 1921 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 1981 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 2041 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 2101 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 2161 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 2221 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 2281 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 2341 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 2401 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 2461 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 2521 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 2581 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 2641 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 2701 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 2761 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 2821 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 2881 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 2941 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 3001 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 3061 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 3121 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 3181 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 3241 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 3301 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 3361 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 3421 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 3481 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 3541 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 3601 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 3661 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 3721 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 3781 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 3841 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 3901 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 3961 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 4021 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 4081 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 4141 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 4201 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 4261 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 4321 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 4381 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 4441 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 4501 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 4561 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc 4621 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat 4681 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca 4741 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg 4801 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa 4861 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa 4921 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata 4981 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag 5041 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc 5101 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta 5161 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg 5221 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt 5281 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca 5341 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag 5401 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa 5461 attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ctgccaccat 5521 ggggaaggtg ctatccaaaa tcttcgggaa caaggaaatg cggatcctca tgttgggcct 5581 ggacgcagcc ggcaagaaca cgatcctgta caagttgaag ctgggccagt cggtgaccac 5641 cattcccact gtgggtttca acgtggagac ggtgacttac aaaaatgtca agttcaacgt 5701 atgggatgtg ggcggccagg acaagatccg gccgctctgg cggcattact acactgggac 5761 ccaaggtctc atcttcgtag tggactgcgc cgaccgcgac cgcatcgatg aggctcgcca 5821 ggagctgcac cgcattatca atgaccggga gatgagggac gccataatcc tcatcttcgc 5881 caacaagcag gacctgcccg atgccatgaa accccacgag atccaggaga aactgggcct 5941 gacccggatt cg //