LOCUS Exported 7876 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: Focal Adhesions, Excitation: 568, Emission: 592. ACCESSION . VERSION . KEYWORDS mApple-Alpha-V-Integrin-25 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7876) TITLE Davidson lab - mApple constructs REFERENCE 2 (bases 1 to 7876) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7876 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 3704..3756 /label=MCS /note="multiple cloning site" regulatory 3766..3775 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3772..4482 /codon_start=1 /product="photostable monomeric derivative of DsRed (Shaner et al., 2008)" /label=mApple /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind 4185..4204 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 4604..4725 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4641..4660) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4695..4714 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4732..5187) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4869..4890) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5081..5100 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5214..5318 /gene="bla" /label=AmpR promoter promoter 5320..5677 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5343..5363) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5528..5663 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5590..5609 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5712..6506 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5766..5785) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6376..6395 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6694..6713) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6738..6785 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7114..7702 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7603..7622 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagtcgac 541 ggtacctcgg cgatggcttt tccgccgcgg cgacggctgc gcctcggtcc ccgcggcctc 601 ccgcttcttc tctcgggact cctgctacct ctgtgccgcg ccttcaacct agacgtggac 661 agtcctgccg agtactctgg ccccgaggga agttacttcg gcttcgccgt ggatttcttc 721 gtgcccagcg cgtcttcccg gatgtttctt ctcgtgggag ctcccaaagc aaacaccacc 781 cagcctggga ttgtggaagg agggcaggtc ctcaaatgtg actggtcttc tacccgccgg 841 tgccagccaa ttgaatttga tgcaacaggc aatagagatt atgccaagga tgatccattg 901 gaatttaagt cccatcagtg gtttggagca tctgtgaggt cgaaacagga taaaattttg 961 gcctgtgccc cattgtacca ttggagaact gagatgaaac aggagcgaga gcctgttgga 1021 acatgctttc ttcaagatgg aacaaagact gttgagtatg ctccatgtag atcacaagat 1081 attgatgctg atggacaggg attttgtcaa ggaggattca gcattgattt tactaaagct 1141 gacagagtac ttcttggtgg tcctggtagc ttttattggc aaggtcagct tatttcggat 1201 caagtggcag aaatcgtatc taaatacgac cccaatgttt acagcatcaa gtataataac 1261 caattagcaa ctcggactgc acaagctatt tttgatgaca gctatttggg ttattctgtg 1321 gctgtcggag atttcaatgg tgatggcata gatgactttg tttcaggagt tccaagagca 1381 gcaaggactt tgggaatggt ttatatttat gatgggaaga acatgtcctc cttatacaat 1441 tttactggcg agcagatggc tgcatatttc ggattttctg tagctgccac tgacattaat 1501 ggagatgatt atgcagatgt gtttattgga gcacctctct tcatggatcg tggctctgat 1561 ggcaaactcc aagaggtggg gcaggtctca gtgtctctac agagagcttc aggagacttc 1621 cagacgacaa agctgaatgg atttgaggtc tttgcacggt ttggcagtgc catagctcct 1681 ttgggagatc tggaccagga tggtttcaat gatattgcaa ttgctgctcc atatgggggt 1741 gaagataaaa aaggaattgt ttatatcttc aatggaagat caacaggctt gaacgcagtc 1801 ccatctcaaa tccttgaagg gcagtgggct gctcgaagca tgccaccaag ctttggctat 1861 tcaatgaaag gagccacaga tatagacaaa aatggatatc cagacttaat tgtaggagct 1921 tttggtgtag atcgagctat cttatacagg gccagaccag ttatcactgt aaatgctggt 1981 cttgaagtgt accctagcat tttaaatcaa gacaataaaa cctgctcact gcctggaaca 2041 gctctcaaag tttcctgttt taatgttagg ttctgcttaa aggcagatgg caaaggagta 2101 cttcccagga aacttaattt ccaggtggaa cttcttttgg ataaactcaa gcaaaaggga 2161 gcaattcgac gagcactgtt tctctacagc aggtccccaa gtcactccaa gaacatgact 2221 atttcaaggg ggggactgat gcagtgtgag gaattgatag cgtatctgcg ggatgaatct 2281 gaatttagag acaaactcac tccaattact atttttatgg aatatcggtt ggattataga 2341 acagctgctg atacaacagg cttgcaaccc attcttaacc agttcacgcc tgctaacatt 2401 agtcgacagg ctcacattct acttgactgt ggtgaagaca atgtctgtaa acccaagctg 2461 gaagtttctg tagatagtga tcaaaagaag atctatattg gggatgacaa ccctctgaca 2521 ttgattgtta aggctcagaa tcaaggagaa ggtgcctacg aagctgagct catcgtttcc 2581 attccactgc aggctgattt catcggggtt gtccgaaaca atgaagcctt agcaagactt 2641 tcctgtgcat ttaagacaga aaaccaaact cgccaggtgg tatgtgacct tggaaaccca 2701 atgaaggctg gaactcaact cttagctggt cttcgtttca gtgtgcacca gcagtcagag 2761 atggatactt ctgtgaaatt tgacttacaa atccaaagct caaatctatt tgacaaagta 2821 agcccagttg tatctcacaa agttgatctt gctgttttag ctgcagttga gataagagga 2881 gtctcgagtc ctgatcatat ctttcttccg attccaaact gggagcacaa ggagaaccct 2941 gagactgaag aagatgttgg gccagttgtt cagcacatct atgagctgag aaacaatggt 3001 ccaagttcat tcagcaaggc aatgctccat cttcagtggc cttacaaata taataataac 3061 actctgttgt atatccttca ttatgatatt gatggaccaa tgaactgcac ttcagatatg 3121 gagatcaacc ctttgagaat taagatctca tctttgcaaa caactgaaaa gaatgacacg 3181 gttgccgggc aaggtgagcg ggaccatctc atcactaagc gggatcttgc cctcagtgaa 3241 ggagatattc acactttggg ttgtggagtt gctcagtgct tgaagattgt ctgccaagtt 3301 gggagattag acagaggaaa gagtgcaatc ttgtacgtaa agtcattact gtggactgag 3361 acttttatga ataaagaaaa tcagaatcat tcctattctc tgaagtcgtc tgcttcattt 3421 aatgtcatag agtttcctta taagaatctt ccaattgagg atatcaccaa ctccacattg 3481 gttaccacta atgtcacctg gggcattcag ccagcgccca tgcctgtgcc tgtgtgggtg 3541 atcattttag cagttctagc aggattgttg ctactggctg ttttggtatt tgtaatgtac 3601 aggatgggct tttttaaacg ggtccggcca cctcaagaag aacaagaaag ggagcagctt 3661 caacctcatg aaaatggtga aggaaactca gaaactccgg gatctcgagc tcaagcttcg 3721 aattctgcag tcgacggtac cgcgggcccg ggatccccac cggtcgccac catggtgagc 3781 aagggcgagg agaataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg 3841 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac 3901 gaggcctttc agaccgctaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg 3961 gacatcctgt cccctcagtt catgtacggc tccaaggtct acattaagca cccagccgac 4021 atccccgact acttcaagct gtccttcccc gagggcttca ggtgggagcg cgtgatgaac 4081 ttcgaggacg gcggcattat tcacgttaac caggactcct ccctgcagga cggcgtgttc 4141 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag 4201 aagaccatgg gctgggaggc ctccgaggag cggatgtacc ccgaggacgg cgccctgaag 4261 agcgagatca agaagaggct gaagctgaag gacggcggcc actacgccgc cgaggtcaag 4321 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacatcgt cgacatcaag 4381 ttggacatcg tgtcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag 4441 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagcggccgc gactctagat 4501 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 4561 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 4621 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 4681 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa 4741 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 4801 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 4861 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 4921 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 4981 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 5041 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 5101 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 5161 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa 5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct 5341 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 5401 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 5461 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 5521 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 5581 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 5641 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag 5701 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 5761 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 5821 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 5881 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 5941 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 6001 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 6061 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 6121 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 6181 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga 6241 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 6301 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 6361 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 6421 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 6481 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 6541 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 6601 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 6661 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga 6721 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt 6781 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc 6841 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg 6901 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta 6961 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 7021 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 7081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 7141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 7201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 7261 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 7321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 7381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 7441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 7501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 7561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 7621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 7681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 7741 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 7801 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc 7861 catatatgga gttccg //