LOCUS Exported 5906 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of untagged ILID CAAX. ACCESSION . VERSION . KEYWORDS ILID-CAAX SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5906) AUTHORS O'Neill PR, Kalyanaraman V, Gautam N TITLE Subcellular optogenetic activation of Cdc42 controls local and distal signaling to drive immune cell migration. JOURNAL Mol Biol Cell. 2016 May 1;27(9):1442-50. doi: 10.1091/mbc.E15-12-0832. Epub 2016 Mar 3. PUBMED 26941336 REFERENCE 2 (bases 1 to 5906) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5906 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(106..126) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 111..440 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 291..426 /label=SV40 ori /note="SV40 origin of replication" primer_bind 353..372 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 507..1301 /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(561..580) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1171..1190 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1475..1596 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1512..1531) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1566..1585 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1645..1661) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1645..1661) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1658..1680) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1669..1685 /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(1693..1723) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1738..1759 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1876..1893) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2047..2635) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2127..2146) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2806..3666) /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 3429..3448 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3667..3771) /gene="bla" /label=AmpR promoter primer_bind complement(3846..3865) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 4037..4416 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4417..4620 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4571..4591 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4665..4684 /label=T7 /note="T7 promoter, forward primer" promoter 4665..4683 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5299..5316) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5305..5529 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5575..97 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5662..5681) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5872..5893 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 61 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 121 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 181 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 241 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 301 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 361 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 421 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 481 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 541 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 601 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 661 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 721 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 781 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 841 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 901 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 961 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 1021 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 1081 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1141 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1201 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1261 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 1321 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 1381 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 1441 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 1501 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 1561 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 1621 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 1681 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 1741 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 1801 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 1861 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 1921 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 1981 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 2041 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 2101 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 2161 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 2221 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 2281 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 2341 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 2401 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 2461 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 2521 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 2581 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 2641 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 2701 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 2761 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 2821 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 2881 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 2941 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 3001 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 3061 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 3121 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 3181 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 3241 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 3301 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 3361 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 3421 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 3481 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 3541 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 3601 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 3661 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 3721 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 3781 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact 3841 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg 3901 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc 3961 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg 4021 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg 4081 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg 4141 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata 4201 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc 4261 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac 4321 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg 4381 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc 4441 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc 4501 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc 4561 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct 4621 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat 4681 agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccatgctg gcaaccacac 4741 tggaacggat cgagaaaaat ttcgtgatta ctgatccgag actgcctgac aacccaatca 4801 tttttgcgag cgattccttc ctgcagctga cagaatattc tcgggaagag atcctggggc 4861 gcaattgccg ttttctgcag ggacccgaga cagaccgtgc cactgttcgg aaaatcagag 4921 atgctattga caaccagact gaagtgaccg ttcagctgat caattatacc aagagcggca 4981 agaagttctg gaacgtgttc cacctgcagc cgatgcgcga ttataagggc gacgtccagt 5041 acttcattgg cgtgcagctg gatggcaccg aacgtcttca tggcgccgct gagcgtgagg 5101 cggtctgcct gatcaaaaag acagcctttc agattgctga ggcagcgaac gacgaaaatt 5161 actttggaag cgggagtggg agcaaaaaga agaaaaagaa gtcaaagaca aagtgtgtaa 5221 ttatgtagga attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg 5281 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 5341 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 5401 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 5461 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 5521 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 5581 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 5641 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 5701 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 5761 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 5821 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 5881 tgttccaaac tggaacaaca ctcaac //