LOCUS Exported 5422 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS GFP-Bcl2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5422) AUTHORS Wang NS, Unkila MT, Reineks EZ, Distelhorst CW TITLE Transient expression of wild-type or mitochondrially targeted Bcl-2 induces apoptosis, whereas transient expression of endoplasmic reticulum-targeted Bcl-2 is protective against Bax-induced cell death. JOURNAL J Biol Chem. 2001 Nov 23. 276(47):44117-28. PUBMED 11546793 REFERENCE 2 (bases 1 to 5422) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5422 /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 605..614 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 611..1327 /codon_start=1 /product="Emerald GFP" /label=EmGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(656..677) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(917..936) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1264..1285 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2217..2338 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2254..2273) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2308..2327 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2345..2800) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2482..2503) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2694..2713 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2827..2931 /gene="bla" /label=AmpR promoter promoter 2933..3290 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2956..2976) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3141..3276 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3203..3222 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3325..4119 /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(3379..3398) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3989..4008 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4307..4326) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4351..4398 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4727..5315 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5216..5235 /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 agccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt 661 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga 721 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc 781 ctggcccacc ctcgtgacca ccttgaccta cggcgtgcag tgcttcgccc gctaccccga 841 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg 901 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg 961 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat 1021 cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca ccgccgacaa 1081 gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt 1141 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc 1201 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga 1261 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct 1321 gtacaagctc agatctatgg cgcacgctgg gagaacgggg tacgataacc gggagatagt 1381 gatgaagtac atccattata agctgtcgca gaggggctac gagtgggatg cgggagatgt 1441 gggcgccgcg cccccggggg ccgcccccgc accgggcatc ttctcctccc agcccgggca 1501 cacgccccat ccagccgcat cccgcgaccc ggtcgccagg acctcgccgc tgcagacccc 1561 ggctgccccc ggcgccgccg cggggcctgc gctcagcccg gtgccacctg tggtccacct 1621 gaccctccgc caggccggcg acgacttctc ccgccgctac cgccgcgact tcgccgagat 1681 gtccagccag ctgcacctga cgcccttcac cgcgcgggga cgctttgcca cggtggtgga 1741 ggagctcttc agggacgggg tgaactgggg gaggattgtg gccttctttg agttcggtgg 1801 ggtcatgtgt gtggagagcg tcaaccggga gatgtcgccc ctggtggaca acatcgccct 1861 gtggatgact gagtacctga accggcacct gcacacctgg atccaggata acggaggctg 1921 ggatgccttt gtggaactgt acggccccag catgcggcct ctgtttgatt tctcctggct 1981 gtctctgaag actctgctca gtttggccct ggtgggagct tgcatcaccc tgggtgccta 2041 tctgagccac aagtgagaat tctgcagtcg acggtaccgc gggcccggga tccaccggat 2101 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa 2161 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact 2221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 2281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac 2341 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc 2401 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 2461 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 2521 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 2581 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 2641 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 2701 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 2761 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga 2821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 2881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg 2941 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa 3301 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3361 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3421 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 3481 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg 3541 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 3601 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 3661 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 3721 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 3781 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 3841 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 3901 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 3961 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4021 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4081 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4141 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4201 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4261 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg 4321 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg 4381 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata 4441 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc 4501 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca 4561 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 4621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 4681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 4741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 4801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 4861 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 4921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 4981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 5041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 5101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 5161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 5221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 5281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 5341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 5401 ctgtggataa ccgtattacc gc //