LOCUS Exported 7002 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Localization: Centrosomes, C1 Cloning Vector, Excitation:487, Emission:509. ACCESSION . VERSION . KEYWORDS mEmerald-Fidgetin-C-18 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7002) TITLE Michael Davidson Unpublished REFERENCE 2 (bases 1 to 7002) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7002 /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" regulatory 547..556 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 553..1269 /codon_start=1 /product="brightly fluorescent monomeric variant of GFP" /label=mEmerald /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(598..619) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(859..878) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1206..1227 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3730..3851 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3767..3786) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3821..3840 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3858..4313) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3995..4016) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4207..4226 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4340..4444 /gene="bla" /label=AmpR promoter promoter 4446..4803 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4469..4489) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4654..4789 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4716..4735 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4838..5632 /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(4892..4911) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5502..5521 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5820..5839) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5864..5911 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6240..6828 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6729..6748 /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 gctagcgcta 541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 721 ccctggccca ccctcgtgac caccttgacc tacggcgtgc agtgcttcgc ccgctacccc 781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 961 atcctggggc acaagctgga gtacaactac aacagccaca aggtctatat caccgccgac 1021 aagcagaaga acggcatcaa ggtgaacttc aagacccgcc acaacatcga ggacggcagc 1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1141 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc 1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1261 ctgtacaagt ccggactcag atctcgagga gggtccagcg gcggtggatc tgggtccagc 1321 ggaatcagta gcaccagtgt ttatggcttg aagatgcagt ggacgccgga gcatgcccag 1381 tggccagaac agcactttga tatcacctcg actactcggt ctcccgccca caaagttgaa 1441 gcctatagag gtcatttgca acgcacctac cagtatgcct gggcgaatga tgacatatct 1501 gctctgactg catccaacct tctgaaaaaa tatgcagaga aatattctgg cattttggaa 1561 ggtcccgtag accgacctgt actcagcaat tattcagaca caccgtcggg actagtgaat 1621 ggtcggaaaa atgatagcga accctggcag tcttccttga attcagaagc cgtttatcct 1681 atgaactgtg ttccggatgt catcaccgcc agcaaagctg gtgtcagttc agccctccct 1741 ccagtagatg tctcggcaag tataggaagc tcccctgggg tggccagcaa cctgacagaa 1801 ccaagttact caagtagtac ctgtggaagc cacacagtac ctagtcttca tgcggggctc 1861 ccgtctcagg aatatgctcc aggatacaac ggatcgtatt tgcattctac ttacagcagc 1921 cagcccacac ctgcacttcc ttcgcctcat ccatccccgt tgcatagctc tgggctcctg 1981 cagccaccac cccctcctcc tcctccaccc gccctggtcc caggctacaa tgggacttcc 2041 aatctctcca gttacagcta tccctctgct agctatcctc ctcagactgc cgtggggtct 2101 gggtacagcc ctggtggagc accccctcct ccttccgcct acctgccttc aggaattcct 2161 gcgcccaccc ctttgccccc cactactgtt cctggctaca cctaccaagg tcatggtttg 2221 acacccatcg caccctctgc tctgaccaac agttcggcaa gttctctcaa aaggaaagct 2281 ttctatatgg cagggcaagg agacatggac tccagttacg gaaattacag ctatggccaa 2341 cagagatcaa cgcagagtcc tatgtaccga atgcccgaca acagcatttc aaactcgaat 2401 cgggggaatg gctttgacag aaatgctgaa acatcatcct tagcatttaa gccaacaaag 2461 cagctgatgc cctctgagca gcaaaggaaa ttcagcagcc agtccagtag ggctctgacc 2521 cctccttcct atagtactgc taaaaattca ttgggatcaa gatcgagtga atcctttggg 2581 aagtatacgt cgccagtcat gagtgagcac ggggacgacc atcggcagct cctcgctcat 2641 ccaatacaag gtccgggact ccgtgcagct acctcatcca accactccgt ggacgagcaa 2701 ctgaagaata ctgacacgca cctcattgac ctggtcacca atgagatcat cacccaagga 2761 cctcctgtgg actggagcga catagctggt cttgacctgg tgaaggcggt tattaaggag 2821 gaggttttgt ggccagtgtt gaggtcagat gcgttcagtg ggctgacagc cttacctagg 2881 agcatccttc tctttggacc tcggggtaca ggcaaaacac tattgggcag atgcatagcc 2941 agtcagctgg gggccacgtt tttcaaaatt gctggttccg gactcgttgc aaagtggcta 3001 ggagaagcgg agaaaattat ccacgcctct tttcttgtgg ccaggtgtcg ccagccctcg 3061 gtgatttttg ttagtgacat tgacatgctt ctctcctctc aagtgagtga ggaacacagt 3121 ccagtgagtc ggatgagaac cgaatttctg atgcagctgg acactgtact gacttcggct 3181 gaggaccaaa tcgtagtcat ttgtgccacc agtaaaccag aagaaataga cgaatctctc 3241 cggaggtact tcatgaaaag acttttaatc ccacttcctg acagcacagc gaggcaccag 3301 ataatagtac aactgctctc acagcacaat tactgtctca atgacaagga gtttgcactg 3361 ctcgtccagc gcacagaagg cttttctgga ctagacgtgg ctcatttgtg ccaggaagca 3421 gcagtgggtc ctctccatgc tatgccagct acagaccttt cagccattat gcccagccag 3481 ttaaggccca ttacatatca agactttgaa aatgcattct gcaagattca gcctagcata 3541 tctcaaaaag agcttgatat gtatgttgaa tggaacaaaa tgtttggttg cagtcagtga 3601 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta 3661 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 3721 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 3781 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 3841 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt 3901 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc 3961 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt 4021 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact 4081 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg 4141 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag 4201 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac 4261 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg 4321 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 4381 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 4441 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc 4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4621 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 4681 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 4741 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 4801 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 4861 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 4921 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 4981 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc 5041 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 5101 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 5161 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 5221 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 5281 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 5341 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca 5401 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 5461 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 5521 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 5581 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 5641 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 5701 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 5761 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact 5821 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag 5881 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg 5941 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt 6001 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg 6061 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat 6121 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 6181 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 6241 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 6301 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 6361 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 6421 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 6481 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 6541 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 6601 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 6661 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 6721 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 6781 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 6841 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 6901 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc 6961 aattacgggg tcattagttc atagcccata tatggagttc cg //