LOCUS Exported 4974 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMito-iRFP670 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4974) AUTHORS Shcherbakova DM, Verkhusha VV TITLE Near-infrared fluorescent proteins for multicolor in vivo imaging. JOURNAL Nat Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521. PUBMED 23770755 REFERENCE 2 (bases 1 to 4974) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4974 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 597..671 /codon_start=1 /product="mitochondrial presequence of human cytochrome c oxidase subunit VIII (Rizutto et al., 1989)" /label=COX8 presequence /translation="MSVLTPLLLRGLTGSARRLPVPRAK" regulatory 699..708 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 705..1640 /codon_start=1 /product="near-infrared fluorescent protein with an emission peak at 670 nm, engineered from a bacterial phytochrome (Shcherbakova and Verkhusha, 2013)" /label=iRFP670 /note="derived from Rhodopseudomonas palustris RpBphP6" /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE PRAVPMAQRVAAEMFADFLSLHFTAAHHQR" polyA_signal 1762..1883 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1799..1818) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1853..1872 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1890..2345) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2027..2048) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2239..2258 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2372..2476 /gene="bla" /label=AmpR promoter promoter 2478..2835 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2501..2521) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2686..2821 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2748..2767 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2870..3664 /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(2924..2943) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3534..3553 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3852..3871) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3896..3943 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4272..4860 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4761..4780 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgt 601 ccgtcctgac gccgctgctg ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc 661 cgcgcgccaa gatccattcg ttgggggatc caccggtcgc caccatggcg cgtaaggtcg 721 atctcacctc ctgcgatcgc gagccgatcc acatccccgg cagcattcag ccgtgcggct 781 gcctgctagc ctgcgacgcg caggcggtgc ggatcacgcg cattacggaa aatgccggcg 841 cgttctttgg acgcgaaact ccgcgggtcg gtgagctact cgccgattac ttcggcgaga 901 ccgaagccca tgcgctgcgc aacgcactgg cgcagtcctc cgatccaaag cgaccggcgc 961 tgatcttcgg ttggcgcgac ggcctgaccg gccgcacctt cgacatctca ctgcatcgcc 1021 atgacggtac atcgatcatc gagttcgagc ctgcggcggc cgaacaggcc gacaatccgc 1081 tgcggctgac gcggcagatc atcgcgcgca ccaaagaact gaagtcgctc gaagagatgg 1141 ccgcacgggt gccgcgctat ctgcaggcga tgctcggcta tcaccgcgtg atgttgtacc 1201 gcttcgcgga cgacggctcc gggatggtga tcggcgaggc gaagcgcagc gacctcgaga 1261 gctttctcgg tcagcacttt ccggcgtcgc tggtcccgca gcaggcgcgg ctactgtact 1321 tgaagaacgc gatccgcgtg gtctcggatt cgcgcggcat cagcagccgg atcgtgcccg 1381 agcacgacgc ctccggcgcc gcgctcgatc tgtcgttcgc gcacctgcgc agcatctcgc 1441 cctgccatct cgaatttctg cggaacatgg gcgtcagcgc ctcgatgtcg ctgtcgatca 1501 tcattgacgg cacgctatgg ggattgatca tctgtcatca ttacgagccg cgtgccgtgc 1561 cgatggcgca gcgcgtcgcg gccgaaatgt tcgccgactt cttatcgctg cacttcaccg 1621 ccgcccacca ccaacgctaa gcggccgcga ctctagatca taatcagcca taccacattt 1681 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 1741 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 1801 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 1861 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt 1921 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 1981 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 2041 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 2101 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 2161 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 2221 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 2281 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 2341 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 2401 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 2461 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg 2521 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2581 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 2641 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 2701 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 2761 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 2821 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca 2881 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 2941 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 3001 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc 3061 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 3121 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 3181 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 3241 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 3301 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 3361 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct 3421 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 3481 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 3541 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 3601 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 3661 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 3721 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 3781 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg 3841 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 3901 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg 3961 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 4021 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 4081 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat 4141 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 4201 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 4261 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 4321 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 4381 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 4441 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 4501 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 4561 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 4621 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 4681 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 4741 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 4801 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 4861 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 4921 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat //