LOCUS Exported 4731 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: C1 Cloning Vector, Excitation: 505, Emission: 515. ACCESSION . VERSION . KEYWORDS mClover2-C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4731) 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 4731) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4731 /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" regulatory 614..623 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 620..1336 /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(665..686) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 1273..1294 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1337..1402 /label=MCS /note="multiple cloning site" polyA_signal 1526..1647 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1563..1582) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1617..1636 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1654..2109) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1791..1812) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2003..2022 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2136..2240 /gene="bla" /label=AmpR promoter promoter 2242..2599 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2265..2285) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2450..2585 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2512..2531 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2634..3428 /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(2688..2707) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3298..3317 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3616..3635) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3660..3707 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4036..4624 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4525..4544 /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 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtcc gcggcgaggg 721 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 781 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcgtggcct gcttcagccg 841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 901 ccaggagcgc accatctctt tcaaggacga cggtacctac aagacccgcg ccgaggtgaa 961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 1021 cggcaacatc ctggggcaca agctggagta caacttcaac agccactacg tctatatcac 1081 ggccgacaag cagaagaaca gcatcaaggc taacttcaag atccgccaca acgttgagga 1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 1201 gctgctgccc gacaaccact acctgagcca tcagtccaag ctgagcaaag accccaacga 1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 1321 ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga 1381 cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca gccataccac 1441 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca 1501 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata 1561 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 1621 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa 1681 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 1741 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 1801 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 1861 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc 1921 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 1981 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 2041 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac 2101 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 2161 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 2221 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt 2281 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 2341 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 2401 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 2461 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 2521 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg 2581 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg 2641 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 2701 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 2761 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg 2821 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 2881 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 2941 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3001 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3061 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3121 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg 3181 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3241 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 3301 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 3361 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 3421 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 3481 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 3541 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 3601 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac 3661 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt 3721 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc 3781 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc 3841 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat 3901 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 3961 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 4021 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 4081 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 4141 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 4201 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 4261 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 4321 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 4381 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 4441 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 4501 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 4561 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 4621 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 4681 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg c //