LOCUS Exported 9030 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses scarlet tagged Kif5a. ACCESSION . VERSION . KEYWORDS pCAG-Scarlet-Kif5a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9030) TITLE Koller Lab plasmids REFERENCE 2 (bases 1 to 9030) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9030 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..657 /label=chicken beta-actin promoter intron 658..1674 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1598..1620 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1682..1701 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1749..1758 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1755..2450 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(1888..1906) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2156..2175 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 5707..5828 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5744..5763) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5798..5817 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5835..6290) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5972..5993) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6184..6203 /label=F1ori-R /note="F1 origin, reverse primer" promoter 6317..6421 /gene="bla" /label=AmpR promoter promoter 6423..6780 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(6446..6466) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6631..6766 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6693..6712 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6815..7609 /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(6869..6888) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7479..7498 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7797..7816) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7841..7888 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 8217..8805 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8706..8725 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8959..8976 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct 421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga 481 tgggggcggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc 541 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct 601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga 661 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc 721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg 781 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc 841 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg 901 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc 961 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc 1081 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc 1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag 1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc 1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc 1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg 1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgt acgctagcgc 1741 taccggtcgc caccatggtg agcaagggcg aggcagtgat caaggagttc atgcggttca 1801 aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg 1861 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc 1921 ccttctcctg ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcaccaagc 1981 accccgccga catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc 2041 gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg 2101 acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg 2161 taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg 2221 gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg 2281 cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg 2341 tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg 2401 aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag tccggactca 2461 gatctcgact gaccatggcg gagactaaca acgaatgcag catcaaggtg ctttgccgat 2521 ttcggcccct gaaccaggcc gagattctgc ggggggacaa gttcatcccc attttccaag 2581 gggacgacag cgtcattatt gggggaaagc catatgtctt tgaccgcgtc ttccccccaa 2641 acaccactca ggagcaggtt taccacgcct gtgccatgca gatcgtcaaa gacgtccttg 2701 ctggttacaa tggcacaatc ttcgcttatg gacagacatc ctcagggaaa acgcatacca 2761 tggaggggaa gctgcacgac cctcagctga tgggcatcat tccccggatc gctcgagaca 2821 tcttcaacca catctactcc atggatgaga accttgaatt ccacattaag gtatcttact 2881 tcgagattta cctggataag atccgtgacc ttttggatgt gaccaagacg aacctgtccg 2941 tgcatgagga caaaaaccgg gtgccgtttg tcaagggttg taccgaacgc tttgtgtcca 3001 gcccagagga gattctggat gtgatcgatg aggggaagtc caaccgtcac gtagctgtca 3061 ccaacatgaa cgagcacagt tctcggagcc acagcatctt cctcatcaac atcaagcagg 3121 agaacgtaga gaccgagcag aagctcagcg ggaagctgta cctcgtggat ctggccggaa 3181 gcgagaaggt cagcaagaca ggggcagagg gagccgttct ggacgaggca aagaatatca 3241 acaagtcgct gtcggccctg gggaacgtga tctctgcact ggcagagggc accaaaagct 3301 acgtgccgta ccgcgacagc aaaatgacga ggattctcca ggactctctg ggagggaact 3361 gcaggactac catgttcatc tgctgctcgc cgtccagcta caatgacgca gagaccaagt 3421 ccacgctcat gtttgggcag cgggcgaaga ccatcaagaa cactgcctca gtgaatctgg 3481 agctgactgc tgagcagtgg aagaagaagt atgagaagga gaaggagaag accaagaccc 3541 agaaggagac aattgcgaag ctagaggctg agcttagccg gtggcgcaat ggagagaatg 3601 tgcctgagac tgagcgcctg gctggagagg actcagctct gggagctgag ctctgcgagg 3661 agacccctgt gaatgacaac tcatccattg tggtacgcat cgcacctgag gaaaggcaga 3721 aatatgagga agagatccgc cgtctctaca agcagcttga tgacaaggat gatgagatca 3781 accagcagag ccagctcatt gagaagctga agcagcagat gctggaccag gaagagctgc 3841 tcgtgtccac tcggggagac aacgagaagg tccagcggga gcttagccac ctgcagtccg 3901 agaacgatgc tgcgaaggac gaggtgaagg aagtgctgca ggccctagag gagctggcgg 3961 tcaactacga ccagaagtcc caggaggtgg aggagaagag ccagcagaac cagctgctgg 4021 tggacgagct gtcccagaaa gtggccacca tgctgtccct ggagtccgag ctacagcggc 4081 tccaggaggt cagtggacac cagcgaaagc ggatcgctga ggtgctgaat gggctgatga 4141 gggacctgag tgagttcagt gtcatcgtgg gcaacggcga gattaagctg cccgtggaga 4201 tcagtggggc catcgaggag gagttcacgg tggcccggct ctacatcagc aagatcaagt 4261 cggaggtgaa gtccgtggtt aagcgatgtc ggcagctgga gaacctccag gtggagtgtc 4321 atcgcaagat ggaggtgacc ggtagggagc tgtcatcttg ccaactgctc atctcacagc 4381 atgaggccaa gatccgttca ctcacggagt acatgcagac tgtggagttg aagaaacggc 4441 acctggaaga gtcctacgac tccctgagcg atgagcttgc caggctccag gcgcacgaaa 4501 ctgtacacga ggtagctctg aaagacaagg agccagacac acaggacgcg gaggaggtga 4561 agaaggccct ggaactacag atggagaatc accgtgaggc ccatcaccgg cagctggccc 4621 gcctccggga tgagattaat gagaaacaga aaaccattga tgagctgaaa gacctgaacc 4681 agaagctcca gttagagctg gagaagcttc aggccgacta tgagaggctg aagaatgaag 4741 agaatgagaa gagcgccaag ctccaggagc tgacatttct gtatgagcga catgagcagt 4801 ccaagcagga cctcaagggg ctggaggaga cagttgcccg tgaactccag accctccaca 4861 accttcgcaa gctgttcgtt caagacgtca cgactcgagt caagaaaagt gcagaaatgg 4921 agcccgagga cagtgggggg attcattccc aaaagcagaa gatctccttt cttgagaaca 4981 acctggaaca gcttacaaag gttcacaaac agctggtacg tgacaatgca gatctgcgtt 5041 gtgagcttcc taaattggaa aaacgacttc gggctacggc tgagagagtt aaggccctgg 5101 agggtgcact gaaggaggcc aaggagggtg ctatgaagga caagcgtaga taccagcagg 5161 aggtggaccg catcaaggaa gccgtgcggt acaagagctc cggcaagcgg ggccattctg 5221 cccagatcgc taagcctgtg aggcctggcc actatcctgc ctcctcaccc accaacccct 5281 acggcacccg gagccccgag tgtatcagct acaccaacaa cctcttccag aactaccaga 5341 acctgcacct gcaggctgcg cctagctcca cttcagatat gtactttgcc agtagcggag 5401 ccacatctgt tgcccccttg gcttcctacc agaaggccaa catggacaat ggaaatgcca 5461 cagatatcaa cgacaacagg agtgacctgc cgtgtggcta tgaggctgag gaccaggcca 5521 agcttttccc tctccaccaa gagacagcag ccagctaact cccaaccctg gccacatgga 5581 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt 5641 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 5701 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 5761 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 5821 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 5881 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 5941 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 6001 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 6061 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 6121 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 6181 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 6241 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 6301 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 6361 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 6421 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 6481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 6541 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 6601 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6661 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6721 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 6781 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 6841 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 6901 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 6961 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 7021 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 7081 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 7141 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 7201 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 7261 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 7321 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 7381 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 7441 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 7501 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 7561 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 7621 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 7681 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 7741 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 7801 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 7861 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 7921 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 7981 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 8041 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 8101 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 8161 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 8221 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 8281 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 8341 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 8401 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 8461 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 8521 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 8581 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 8641 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 8701 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 8761 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 8821 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 8881 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 8941 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 9001 ggccggccaa ggtggccacg cgtacatgtc //