LOCUS Exported 4968 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGFH-9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4968) AUTHORS Mary C, Scherrer A, Huck L, Lakkaraju AK, Thomas Y, Johnson AE, Strub K TITLE Residues in SRP9/14 essential for elongation arrest activity of the signal recognition particle define a positively charged functional domain on one side of the protein. JOURNAL RNA. 2010 May;16(5):969-79. Epub 2010 Mar 26. PUBMED 20348448 REFERENCE 2 (bases 1 to 4968) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4968 /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" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 613..1329 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1756..1877 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1793..1812) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1847..1866 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1884..2339) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2021..2042) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2233..2252 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2366..2470 /gene="bla" /label=AmpR promoter promoter 2472..2829 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2495..2515) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2680..2815 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2742..2761 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2864..3658 /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(2918..2937) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3528..3547 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3846..3865) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3890..3937 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4266..4854 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4755..4774 /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctatgcc gcagtaccag 1381 acctgggagg agttcagccg cgctgccgag aagctttacc tcgctgaccc tatgaaggca 1441 cgtgtggttc tcaaatatag gcattctgat gggaacttgt gtgttaaagt aacagatgat 1501 ttagtttgtt tggtgtataa aacagaccaa gctcaagatg taaagaagat tgagaaattc 1561 cacagtcaac taatgcgact tatggtagcc aaggaagccc gcaatgttac catggaaact 1621 gagtgaggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag 1681 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat 1741 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc 1801 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 1861 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt 1921 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta 1981 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc 2041 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg 2101 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact 2161 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt 2221 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc 2281 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc 2341 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 2401 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 2461 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga 2521 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2581 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 2641 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 2701 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 2761 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 2821 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg 2881 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca 2941 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt 3001 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg 3061 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga 3121 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca 3181 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct 3241 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac 3301 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc 3361 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt 3421 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt 3481 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg 3541 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat 3601 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc 3661 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc 3721 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 3781 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg 3841 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa 3901 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag 3961 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct 4021 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc 4081 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa 4141 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 4201 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 4261 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 4321 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 4381 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 4441 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 4501 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 4561 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 4621 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 4681 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 4741 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 4801 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 4861 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 4921 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat //