LOCUS Exported 7097 bp DNA circular SYN 18-OCT-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7097) AUTHORS Institute for Molecular Bioscience TITLE Direct Submission JOURNAL Exported Jun 7, 2022 from SnapGene Viewer 6.0.5 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7097 /mol_type="other DNA" /organism="synthetic DNA construct" 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" misc_feature 591..595 /label=MCS /note="multiple cloning site of fluorescent protein plasmids" mRNA 596..1378 /product="interleukin-2 receptor alpha-subunit" misc_feature 1394..2377 /label=AH /note="Anillin homology domain" CDS 2396..2449 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 2468..3175 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind 2468..2491 /label=Primer 2 primer_bind complement(3149..3175) /label=Primer 1 CDS 3182..3763 /codon_start=1 /product="constitutively active mutant of human Rho family small GTPase RhoA" /label=RhoA (Q63L) /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD IEVDGKQVELALWDTAGLEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT KDGVREVFEMATRAALQARRGKKKSGCLVL" polyA_signal 3885..4006 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(4013..4468) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4495..4599 /gene="bla" /label=AmpR promoter promoter 4601..4958 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4809..4944 /label=SV40 ori /note="SV40 origin of replication" CDS 4993..5787 /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)" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" misc_feature 6019..6066 /label=HSV TK poly(A) signal /note="herpesvirus thymidine kinase polyadenylation signal" rep_origin 6395..6983 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" 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 gctagatgga 601 ttcatacctg ctgatgtggg gactgctcac gttcatcatg gtgcctggct gccaggcaga 661 gctctgtgac gatgacccgc cagagatccc acacgccaca ttcaaagcca tggcctacaa 721 ggaaggaacc atgttgaact gtgaatgcaa gagaggtttc cgcagaataa aaagcgggtc 781 actctatatg ctctgtacag gaaactctag ccactcgtcc tgggacaacc aatgtcaatg 841 cacaagctct gccactcgga acacaacgaa acaagtgaca cctcaacctg aagaacagaa 901 agaaaggaaa accacagaaa tgcaaagtcc aatgcagcca gtggaccaag cgagccttcc 961 aggtcactgc agggaacctc caccatggga aaatgaagcc acagagagaa tttatcattt 1021 cgtggtgggg cagatggttt attatcagtg cgtccaggga tacagggctc tacacagagg 1081 tcctgctgag agcgtctgca aaatgaccca cgggaagaca aggtggaccc agccccagct 1141 catatgcaca ggtgaaatgg agaccagtca gtttccaggt gaagagaagc ctcaggcaag 1201 ccccgaaggc cgtcctgaga gtgagacttc ctgcctcgtc acaacaacag attttcaaat 1261 acagacagaa atggctgcaa ccatggagac gtccatattt acaacagagt accaggtagc 1321 agtggccggc tgtgttttcc tgctgatcag cgtcctcctc ctgagtgggc tcacctggaa 1381 gggaaaattc caaagaactc gtgtccctcg agctgaatct ggtgatagcc ttggttctga 1441 agatcgtgat cttctttaca gcattgatgc atatagatct caaagattca aagaaacaga 1501 acgtccatca ataaagcagg tgattgttcg gaaggaagat gttacttcaa aactggatga 1561 aaaaaataat gcctttcctt gtcaagttaa tatcaaacag aaaatgcagg aactcaataa 1621 cgaaataaat atgcaacaga cagtgatcta tcaagctagc caggatctta actgctgtgt 1681 tgatgaagaa catggaaaag ggtccctaga agaagctaaa gcagaaagac ttcttctaat 1741 tgcaactggg aagagaacac ttttgattga tgaattgaat aaattgaaga acgaaggacc 1801 tcagaggaag aataaggcta gtccccaaag tgaatttatg ccatccaaag gatcagttac 1861 tttgtcagaa atccgcttgc ctctaaaagc agattttgtc tgcagtacgg ttcagaaacc 1921 agatgcagca aattactatt acttaattat actaaaagca ggagctgaaa atatggtagc 1981 cacaccatta gcaagtactt caaactctct taacggtgat gctctgacat tcactactac 2041 atttactctg caagatgtat ccaatgactt tgaaataaat attgaagttt acagcttggt 2101 gcaaaagaaa gatccctcag gccttgataa gaagaaaaaa acatccaagt ccaaggctat 2161 tactccaaag cgactcctca catctataac cacaaaaagc aacattcatt cttcagtcat 2221 ggccagtcca ggaggtctta gtgctgtgcg aaccagcaac ttcgcccttg ttggatctta 2281 cacattatca ttgtcttcag taggaaatac taagtttgtt ctggacaagg tccccttttt 2341 atcttctttg gaaggtcata tttatttaaa aataaaatgt caagtgcgac cggtcgaggg 2401 caggggaagc ctgctcacct gcggcgacgt ggaggaaaac cctggccccg ccccaggatc 2461 caagcttatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg 2521 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg 2581 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc 2641 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt 2701 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg 2761 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct 2821 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg 2881 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga 2941 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta 3001 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta 3061 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca 3121 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggaatt 3181 catggctgcc atccggaaga aactggtgat tgttggtgat ggagcctgtg gaaagacatg 3241 cttgctcata gtcttcagca aggaccagtt cccagaggtg tatgtgccca cagtgtttga 3301 gaactatgtg gcagatatcg aggtggatgg aaagcaggta gagttggctt tgtgggacac 3361 agctgggctg gaagattatg atcgcctgag gcccctctcc tacccagata ccgatgttat 3421 actgatgtgt ttttccatcg acagccctga tagtttagaa aacatcccag aaaagtggac 3481 cccagaagtc aagcatttct gtcccaacgt gcccatcatc ctggttggga ataagaagga 3541 tcttcggaat gatgagcaca caaggcggga gctagccaag atgaagcagg agccggtgaa 3601 acctgaagaa ggcagagata tggcaaacag gattggcgct tttgggtaca tggagtgttc 3661 agcaaagacc aaagatggag tgagagaggt ttttgaaatg gctacgagag ctgctctgca 3721 agctagacgt gggaagaaaa aatctggttg ccttgtcttg tgagcggccg cgactctaga 3781 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc 3841 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag 3901 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 3961 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta 4021 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac 4081 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg 4141 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa 4201 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt 4261 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt 4321 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga 4381 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc 4441 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga 4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 4561 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc 4621 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 4681 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 4741 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 4801 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 4861 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt 4921 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga 4981 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 5041 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 5101 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 5161 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 5221 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 5281 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 5341 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 5401 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 5461 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 5521 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 5581 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 5641 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 5701 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 5761 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 5821 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 5881 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 5941 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg 6001 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt 6061 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac 6121 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg 6181 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt 6241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 6301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 6541 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 7021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 7081 gtattaccgc catgcat //