LOCUS Exported 7014 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTfR-PATagRFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7014) AUTHORS Subach FV, Patterson GH, Renz M, Lippincott-Schwartz J, Verkhusha VV TITLE Bright monomeric photoactivatable red fluorescent protein for two-color super-resolution sptPALM of live cells. JOURNAL J Am Chem Soc. 2010 May 12;132(18):6481-91. PUBMED 20394363 REFERENCE 2 (bases 1 to 7014) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7014 /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 complement(1565..1576) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" regulatory 2960..2969 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2984..3679 /codon_start=1 /product="photoactivatable variant of the monomeric red fluorescent protein TagRFP (Subach et al., 2010)" /label=PA-TagRFP /note="mammalian codon-optimized" /translation="ELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKVVE GGPLPFAFDILATSFMYGSSTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTATQ DTSLQDGCLIYNVKIRGVNFPSNGPVMKKKTLGWEPSTEKLKPADGGLEGRVDMALKLV GGGHLICNFKTTYRSKKPAKNLKMPGVYYVDRRLEIIKEADKETYWEQHEVAVARYSDL PSKLGHKLN" polyA_signal 3802..3923 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3839..3858) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3893..3912 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3930..4385) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4067..4088) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4279..4298 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4412..4516 /gene="bla" /label=AmpR promoter promoter 4518..4875 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4541..4561) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4726..4861 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4788..4807 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4910..5704 /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(4964..4983) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5574..5593 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5892..5911) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5936..5983 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6312..6900 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6801..6820 /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 gctagcgcta 601 ccggactcag atctcgagcg ccaccatgat ggatcaagct agatcagcat tctctaactt 661 gtttggtgga gaaccattgt catatacccg gttcagcctg gctcggcaag tagatggcga 721 taacagtcat gtggagatga aacttgctgt agatgaagaa gaaaatgctg acaataacac 781 aaaggccaat gtcacaaaac caaaaaggtg tagtggaagt atctgctatg ggactattgc 841 tgtgatcgtc tttttcttga ttggatttat gattggctac ttgggctatt gtaaaggggt 901 agaaccaaaa actgagtgtg agagactggc aggaaccgag tctccagtga gggaggagcc 961 aggagaggac ttccctgcag cacgtcgctt atattgggat gacctgaaga gaaagttgtc 1021 ggagaaactg gacagcacag acttcaccag caccatcaag ctgctgaatg aaaattcata 1081 tgtccctcgt gaggctggat ctcaaaaaga tgaaaatctt gcgttgtatg ttgaaaatca 1141 atttcgtgaa tttaaactca gcaaagtctg gcgtgatcaa cattttgtta agattcaggt 1201 caaagacagc gctcaaaact cggtgatcat agttgataag aacggtagac ttgtttacct 1261 ggtggagaat cctgggggtt atgtggcgta tagtaaggct gcaacagtta ctggtaaact 1321 ggtccatgct aattttggta ctaaaaaaga ttttgaggat ttatacactc ctgtgaatgg 1381 atctatagtg attgtcagag cagggaaaat cacctttgca gaaaaggttg caaatgctga 1441 aagcttaaat gcaattggtg tgttgatata catggaccag actaaatttc ccattgttaa 1501 cgcagaactt tcattctttg gacatgctca tctggggaca ggtgaccctt acacacctgg 1561 attcccttcc ttcaatcaca ctcagtttcc accatctcgg tcatcaggat tgcctaatat 1621 acctgtccag acaatctcca gagctgctgc agaaaagctg tttgggaata tggaaggaga 1681 ctgtccctct gactggaaaa cagactctac atgtaggatg gtaacctcag aaagcaagaa 1741 tgtgaagctc actgtgagca atgtgctgaa agagataaaa attcttaaca tctttggagt 1801 tattaaaggc tttgtagaac cagatcacta tgttgtagtt ggggcccaga gagatgcatg 1861 gggccctgga gctgcaaaat ccggtgtagg cacagctctc ctattgaaac ttgcccagat 1921 gttctcagat atggtcttaa aagatgggtt tcagcccagc agaagcatta tctttgccag 1981 ttggagtgct ggagactttg gatcggttgg tgccactgaa tggctagagg gatacctttc 2041 gtccctgcat ttaaaggctt tcacttatat taatctggat aaagcggttc ttggtaccag 2101 caacttcaag gtttctgcca gcccactgtt gtatacgctt attgagaaaa caatgcaaaa 2161 tgtgaagcat ccggttactg ggcaatttct atatcaggac agcaactggg ccagcaaagt 2221 tgagaaactc actttagaca atgctgcttt ccctttcctt gcatattctg gaatcccagc 2281 agtttctttc tgtttttgcg aggacacaga ttatccttat ttgggtacca ccatggacac 2341 ctataaggaa ctgattgaga ggattcctga gttgaacaaa gtggcacgag cagctgcaga 2401 ggtcgctggt cagttcgtga ttaaactaac ccatgatgtt gaattgaacc tggactatga 2461 gaggtacaac agccaactgc tttcatttgt gagggatctg aaccaataca gagcagacat 2521 aaaggaaatg ggcctgagtt tacagtggct gtattctgct cgtggagact tcttccgtgc 2581 tacttccaga ctaacaacag atttcgggaa tgctgagaaa acagacagat ttgtcatgaa 2641 gaaactcaat gatcgtgtca tgagagtgga gtatcacttc ctctctccct acgtatctcc 2701 aaaagagtct cctttccgac atgtcttctg gggctccggc tctcacacgc tgccagcttt 2761 actggagaac ttgaaactgc gtaaacaaaa taacggtgct tttaatgaaa cgctgttcag 2821 aaaccagttg gctctagcta cttggactat tcagggagct gcaaatgccc tctctggtga 2881 cgtttgggac attgacaatg agttttcaga attcggctcc accggctcca ccggctccac 2941 cggcgcggat ccaccggtcg ccaccatggt gtctaagggc gaagagctga ttaaggagaa 3001 catgcacatg aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc 3061 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg 3121 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcagcac 3181 cttcatcaac cacacccagg gcatccccga cttctggaag cagtccttcc ctgagggctt 3241 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac 3301 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc 3361 caacggccct gtgatgaaga agaaaacact cggctgggaa ccgagcaccg agaaactgaa 3421 acccgctgac ggcggcctgg aaggcagggt cgacatggcc ctgaagctcg tgggcggggg 3481 ccacctgatc tgcaacttca agaccacata cagatccaag aaacccgcta agaacctcaa 3541 gatgcccggc gtctactatg tggaccgcag actggaaatc atcaaggagg ccgacaaaga 3601 gacctactgg gagcagcacg aggtggctgt ggccagatac tccgacctcc ctagcaaact 3661 ggggcacaag cttaattaaa gcggccgcga ctctagatca taatcagcca taccacattt 3721 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 3781 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 3841 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 3901 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt 3961 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 4021 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 4081 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 4141 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 4201 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 4261 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 4321 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 4381 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 4441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 4501 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg 4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 4621 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4681 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 4741 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 4801 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 4861 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca 4921 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 4981 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 5041 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc 5101 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 5161 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 5221 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 5281 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 5341 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 5401 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct 5461 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 5521 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 5581 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 5641 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 5701 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 5761 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 5821 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg 5881 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 5941 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg 6001 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 6061 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 6121 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat 6181 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 6241 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 6301 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 6361 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 6421 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 6481 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 6541 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 6601 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 6661 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 6721 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 6781 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 6841 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 6901 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 6961 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat //