LOCUS Exported 5824 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: Membrane Scaffold, Excitation: 434, Emission: 474. ACCESSION . VERSION . KEYWORDS mTurquoise-Sequestosome1-C-18 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5824) TITLE Davidson Cyan Fluorescent Proteins REFERENCE 2 (bases 1 to 5824) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5824 /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="enhanced monomeric variant of CFP (Goedhart et al., 2010)" /label=mTurquoise /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL 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 2552..2673 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2589..2608) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2643..2662 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2680..3135) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2817..2838) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3029..3048 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3162..3266 /gene="bla" /label=AmpR promoter promoter 3268..3625 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3291..3311) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3476..3611 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3538..3557 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3660..4454 /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(3714..3733) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4324..4343 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4642..4661) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4686..4733 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5062..5650 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5551..5570 /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 caccctgtcc tggggcgtgc 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 atcagcgaca acgtctatat caccgccgac 1021 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc 1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1141 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc 1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1261 ctgtacaagt ccggactcag atctcgagag agcggaggta gcgcatccgg cggaagcgga 1321 ggagccatgt cctacgtgaa ggatgacatc ttccgaatct acattaaaga gaaaaaagag 1381 tgccggcggg accaccgccc accgtgtgct caggaggcgc cccgcaacat ggtgcacccc 1441 aatgtgatct gcgatggctg caatgggcct gtggtaggaa cccgctacaa gtgcagcgtc 1501 tgcccagact acgacttgtg tagcgtctgc gagggaaagg gcttgcaccg ggggcacacc 1561 aagctcgcat tccccagccc cttcgggcac ctgtctgagg gcttctcgca cagccgctgg 1621 ctccggaagg tgaaacacgg acacttcggg tggccaggat gggaaatggg tccaccagga 1681 aactggagcc cacgtcctcc tcgtgcaggg gaggcccgcc ctggccccac ggcagaatca 1741 gcttctggtc catcggagga tccgagtgtg aatttcctga agaacgttgg ggagagtgtg 1801 gcagctgccc ttagccctct gggcattgaa gttgatatcg atgtggagca cggagggaaa 1861 agaagccgcc tgacccccgt ctctccagag agttccagca cagaggagaa gagcagctca 1921 cagccaagca gctgctgctc tgatcccagc aagccgggtg ggaatgttga gggcgccacg 1981 cagtctctgg cggagcagat gagaaagatc gccttggagt ccgaggggcg ccctgaggaa 2041 cagatggagt cggataactg ttcaggagga gatgatgact ggacccatct gtcttcaaaa 2101 gaagtggacc cgtctacagg tgaactccag tccctacaga tgccagaatc cgaagggcca 2161 agctctctgg acccctccca ggagggaccc acagggctga aggaagctgc cttgtaccca 2221 catctcccgc cagaggctga cccgcggctg attgagtccc tctcccagat gctgtccatg 2281 ggcttctctg atgaaggcgg ctggctcacc aggctcctgc agaccaagaa ctatgacatc 2341 ggagcggctc tggacaccat ccagtattca aagcatcccc cgccgttgtg agaattctgc 2401 agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca 2461 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct 2521 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta 2581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 2641 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt 2701 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa 2761 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag 2821 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg 2881 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga 2941 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg 3001 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg 3061 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc 3121 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 3181 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 3241 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg 3301 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3361 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3421 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3481 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3541 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 3601 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca 3661 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 3721 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 3781 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 3841 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 3901 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 3961 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 4021 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 4081 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 4141 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg 4201 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 4261 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 4321 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 4381 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 4441 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct 4501 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 4561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 4621 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc 4681 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg 4741 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca 4801 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg 4861 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact 4921 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 4981 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 5041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 5101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 5161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 5221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 5281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 5341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 5401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 5461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 5521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 5581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 5641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 5701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat 5761 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt 5821 tccg //