LOCUS Exported 4731 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: C1 Cloning Vector, Excitation: 488, Emission: 507. ACCESSION . VERSION . KEYWORDS mEGFP-C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4731) TITLE Michael Davidson Empty Backbones 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 142..445 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 446..649 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 600..620 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 688..697 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 694..1410 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(739..760) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1000..1019) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1347..1368 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1411..1476 /label=MCS /note="multiple cloning site" polyA_signal 1600..1721 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1637..1656) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1691..1710 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1728..2183) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1865..1886) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2077..2096 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2210..2314 /gene="bla" /label=AmpR promoter promoter 2316..2673 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2339..2359) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2524..2659 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2586..2605 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2708..3502 /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(2762..2781) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3372..3391 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3690..3709) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3734..3781 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4110..4698 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4599..4618 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 61 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca 121 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 181 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 241 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 301 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 361 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 421 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg 481 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt 541 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac 601 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa 661 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc 721 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 781 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 841 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg 901 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 961 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 1021 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 1081 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 1141 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 1201 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 1261 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc 1321 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 1381 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc tcaagcttcg 1441 aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata actgatcata 1501 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc 1561 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat 1621 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 1681 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt 1741 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata 1801 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt 1861 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg 1921 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt 1981 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc 2041 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg 2101 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct 2161 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 2221 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 2281 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg 2341 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 2401 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 2461 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 2521 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 2581 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga 2641 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc 2701 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag 2761 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg 2821 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa 2881 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc 2941 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc 3001 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga 3061 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa 3121 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct 3181 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat 3241 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt 3301 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta 3361 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga 3421 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg 3481 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg 3541 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 3601 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 3661 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga 3721 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt 3781 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat 3841 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa 3901 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca 3961 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 4021 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 4081 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 4141 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 4201 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 4261 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 4321 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 4381 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 4441 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 4501 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 4561 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 4621 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 4681 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct g //