LOCUS Exported 4732 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: N1 Cloning Vector, Excitation: 505, Emission: 515. ACCESSION . VERSION . KEYWORDS mClover2-N1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4732) AUTHORS Bajar BT, Wang ES, Lam AJ, Kim BB, Jacobs CL, Howe ES, Davidson MW, Lin MZ, Chu J TITLE Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting. JOURNAL Sci Rep. 2016 Feb 16;6:20889. doi: 10.1038/srep20889. PUBMED 26879144 REFERENCE 2 (bases 1 to 4732) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4732 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 68..371 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 372..575 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 526..546 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 598..678 /label=MCS /note="multiple cloning site" regulatory 680..689 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 686..1405 /codon_start=1 /product="monomeric derivative of the bright green-yellow fluorescent protein Clover" /label=mClover2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNSIK ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(731..752) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 1339..1360 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1527..1648 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1564..1583) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1618..1637 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1655..2110) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1792..1813) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2004..2023 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2137..2241 /gene="bla" /label=AmpR promoter promoter 2243..2600 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2266..2286) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2451..2586 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2513..2532 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2635..3429 /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(2689..2708) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3299..3318 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3617..3636) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3661..3708 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4037..4625 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4526..4545 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct 601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg 661 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 721 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtccgcgg 781 cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg 841 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcg tggcctgctt 901 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 961 ctacgtccag gagcgcacca tctctttcaa ggacgacggt acctacaaga cccgcgccga 1021 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 1081 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaacagcc actacgtcta 1141 tatcacggcc gacaagcaga agaacagcat caaggctaac ttcaagatcc gccacaacgt 1201 tgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 1261 ccccgtgctg ctgcccgaca accactacct gagccatcag tccaagctga gcaaagaccc 1321 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 1381 cggcatggac gagctgtaca agtaagcggc cgcgactcta gatcataatc agccatacca 1441 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 1501 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat 1561 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 1621 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta 1681 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc 1741 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg 1801 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat 1861 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc 1921 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag 1981 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg 2041 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta 2101 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 2161 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 2221 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt 2281 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 2341 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2401 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 2461 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 2521 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 2581 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt 2641 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 2701 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 2761 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac 2821 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 2881 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 2941 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 3001 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 3061 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 3121 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag 3181 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 3241 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 3301 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 3361 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 3421 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 3481 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 3541 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 3601 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 3661 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg 3721 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 3781 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 3841 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga 3901 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 3961 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 4021 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 4081 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 4141 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 4201 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 4261 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 4321 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 4381 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 4441 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 4501 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 4561 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 4621 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 4681 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gc //