LOCUS Exported 4723 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFast-FT-N1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4723) AUTHORS Subach FV, Subach OM, Gundorov IS, Morozova KS, Piatkevich KD, Cuervo AM, Verkhusha VV TITLE Monomeric fluorescent timers that change color from blue to red report on cellular trafficking. JOURNAL Nat Chem Biol. 2009 Feb;5(2):118-26. Epub 2009 Jan 11. PUBMED 19136976 REFERENCE 2 (bases 1 to 4723) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4723 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 54..357 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 358..561 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 512..532 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 584..664 /label=MCS /note="multiple cloning site" regulatory 666..675 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 672..1382 /codon_start=1 /product="blue-to-red fluorescent timer derivative of mCherry (Subach et al., 2009)" /label=Fast-FT /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHVEGSVNGHEFEIEGEGKGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSRAYVKHPADIPDYWKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERS EGRHSTGGMDELYK" primer_bind complement(817..835) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1085..1104 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 1504..1625 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1541..1560) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1595..1614 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1632..2087) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1769..1790) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1981..2000 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2114..2218 /gene="bla" /label=AmpR promoter promoter 2220..2577 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2243..2263) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2428..2563 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2490..2509 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2612..3406 /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(2666..2685) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3276..3295 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3594..3613) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3638..3685 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4014..4602 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4503..4522 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact 601 cagatctcga gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac 661 cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaagagttca 721 tgcgtttcaa agtgcacgtg gagggttccg tgaacggcca cgagttcgag atcgagggcg 781 agggcaaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg 841 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct 901 acgtgaagca tcccgccgac atccccgact actggaagct gtccttcccc gagggcttca 961 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 1021 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggaactaac ttcccctccg 1081 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cggatgtacc 1141 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc 1201 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg 1261 cctacaacgt caatatcaag ttggacatca catcccacaa cgaggactac accatcgtgg 1321 aacagtacga acgctccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt 1381 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct 1441 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 1501 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 1561 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 1621 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 1681 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 1741 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 1801 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 1861 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 1921 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 1981 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 2041 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt 2101 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 2161 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc 2221 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2281 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 2341 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2401 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 2461 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 2521 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat 2581 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 2641 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 2701 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 2761 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct 2821 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 2881 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 2941 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 3001 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 3061 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 3121 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 3181 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 3241 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 3301 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 3361 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 3421 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 3481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 3541 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca 3601 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 3661 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 3721 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 3781 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 3841 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa 3901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 3961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 4021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 4081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 4141 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 4201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 4261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 4321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 4381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 4441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 4501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 4561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 4621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 4681 cctgattctg tggataaccg tattaccgcc atgcattagt tat //