LOCUS Exported 8474 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEGFP-STAG2-wildtype SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8474) AUTHORS Solomon DA, Kim T, Diaz-Martinez LA, Fair J, Elkahloun AG, Harris BT, Toretsky JA, Rosenberg SA, Shukla N, Ladanyi M, Samuels Y, James CD, Yu H, Kim JS, Waldman T TITLE Mutational inactivation of STAG2 causes aneuploidy in human cancer. JOURNAL Science. 2011 Aug 19;333(6045):1039-43. PUBMED 21852505 REFERENCE 2 (bases 1 to 8474) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8474 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 16..319 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 320..523 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 474..494 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 562..571 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 568..1284 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 568..1284 /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(613..634) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(874..893) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1221..1242 /label=EGFP-C /note="EGFP, forward primer" CDS 5014..5037 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" misc_feature 5041..5093 /label=MCS /note="multiple cloning site" polyA_signal 5217..5338 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5254..5273) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5308..5327 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5345..5800) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5482..5503) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5694..5713 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5827..5931 /gene="bla" /label=AmpR promoter promoter 5933..6290 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5956..5976) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6141..6276 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6203..6222 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6325..7119 /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(6379..6398) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6989..7008 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7307..7326) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7351..7398 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7727..8315 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8216..8235 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca 541 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg 601 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 661 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 721 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 781 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 841 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 901 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 961 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc 1021 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac 1081 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac 1141 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac 1201 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact 1261 ctcggcatgg acgagctgta caagtactca gatctcgacg agctcactag tcgcggccgc 1321 atgatagcag ctccagaaat accaactgat tttaatctac tacaggagtc agaaacacat 1381 ttttcttctg acacagattt tgaagatatc gaaggaaaaa accaaaagca aggcaaaggc 1441 aaaacttgta aaaaaggcaa aaagggccca gcagaaaagg gcaaaggtgg aaatggagga 1501 ggaaaacctc cttctggtcc aaaccgaatg aatggtcatc accaacagaa tggagtggaa 1561 aacatgatgt tgtttgaagt tgttaaaatg ggcaagagtg ctatgcagtc ggtggtagat 1621 gattggatag aatcatacaa gcatgaccga gatatagcac ttcttgacct tatcaacttt 1681 tttattcagt gttcaggctg taaaggagtt gtcacagcag aaatgtttag acatatgcag 1741 aactctgaga taattcgaaa aatgactgaa gaattcgatg aggatagtgg agattatcca 1801 cttaccatgg ctggtcctca gtggaagaag ttcaaatcca gtttttgtga attcattggc 1861 gtgttagtac ggcaatgtca atatagtatc atatatgatg agtatatgat ggatacagtc 1921 atttcacttc ttacaggatt gtctgactca caagtcagag catttcgaca tacaagcacc 1981 ctggcagcta tgaagttgat gacagctttg gtgaatgtgg cactaaatct tagcattaat 2041 atggataata cacaaagaca atatgaagca gaacggaata aaatgattgg aaaacgagcc 2101 aatgagaggc tagaactcct gctacaaaag cggaaagagc ttcaggaaaa tcaagatgaa 2161 atagaaaata tgatgaatgc aatatttaaa ggagtgtttg tacatagata ccgtgatgcg 2221 atagctgaaa ttcgagctat ttgcattgaa gagattggca tttggatgaa gatgtatagt 2281 gatgcctttc ttaatgacag ttatttaaaa tatgttggtt ggactatgca tgataagcaa 2341 ggtgaagtaa gactcaaatg tcttactgct ctacaagggc tttattataa caaagagctt 2401 aattccaaac tggaactttt taccagtcgg ttcaaggata gaattgtgtc tatgaccctt 2461 gacaaagaat atgatgttgc agtacaagca ataaaattac tcactcttgt tttacagagt 2521 agtgaagaag ttctcactgc agaagattgt gaaaatgtct atcatctggt ttattcagct 2581 caccggccag tagcagtagc agctggagaa tttctctaca aaaagctctt cagtcgtaga 2641 gatccagagg aggatggaat gatgaaaaga agaggaagac aaggtccaaa tgccaacctt 2701 gttaagacat tggttttttt ctttctagaa agtgagttac atgagcatgc agcatacctt 2761 gtggatagca tgtgggactg tgctactgag ctgctgaaag actgggaatg tatgaatagc 2821 ttgttactgg aagagccact tagtggagag gaagcactaa cagataggca agagagtgct 2881 ctgattgaaa taatgctttg taccattaga caagcggctg aatgtcatcc tcccgtggga 2941 agagggacag gaaaaagggt gcttacagca aaggagaaga agacacagtt ggatgatagg 3001 acaaaaatca ctgagctttt tgccgtggcc cttcctcagt tattagcaaa atactctgta 3061 gatgcagaaa aggtgactaa cttgttgcag ttgcctcagt actttgattt ggaaatatat 3121 accactggac gattagaaaa gcatttggat gccttattgc gacagatccg gaatattgta 3181 gagaagcaca cagatacaga tgttttggaa gcatgttcta aaacttacca tgcactctgt 3241 aatgaagagt tcacaatctt caacagagta gatatttcaa gaagtcaact gatagatgaa 3301 ttggcagata aatttaaccg gcttcttgaa gattttctgc aagagggtga agaacctgat 3361 gaagatgatg catatcaggt attgtcaaca ttgaagagga tcactgcttt tcataatgcc 3421 catgaccttt caaagtggga tttatttgct tgtaattaca aactcttgaa aactggaatc 3481 gaaaatggag acatgcctga gcagattgtt attcacgcac tgcagtgtac tcactatgta 3541 atcctttggc aacttgctaa gataactgaa agcagctcta caaaggagga cttgctgcgt 3601 ttaaagaaac aaatgagagt attttgtcag atatgtcaac attacctgac caacgtgaat 3661 actactgtta aggaacaggc cttcactatt ctgtgtgata ttttgatgat cttcagccat 3721 cagattatgt caggagggcg tgacatgtta gagccattag tgtatacccc tgattcttca 3781 ttgcagtctg agttgctcag ctttattttg gatcatgtct tcattgaaca ggatgatgat 3841 aataatagtg cagatggtca gcaagaggat gaagccagta aaattgaagc tctgcacaag 3901 agaagaaatt tacttgcagc attttgtaag ctaattgtat atactgtggt ggagatgaat 3961 acagctgcag atatcttcaa acagtatatg aagtattata atgactatgg agatatcatc 4021 aaagaaacaa tgagtaaaac aaggcagata gacaaaattc agtgtgctaa gacccttatt 4081 ctcagtctgc aacagctttt taatgaaatg atacaagaaa atggctataa ttttgataga 4141 tcatcctcta catttagtgg cataaaagaa cttgctcgac gttttgcttt aacttttgga 4201 cttgatcagt tgaaaacaag agaagccatt gccatgctac acaaagatgg catagaattt 4261 gcttttaaag agcctaatcc gcaaggggag agccatccac ctttaaattt ggcatttctt 4321 gatattctga gtgaattttc ttctaaacta cttcgacaag acaaaagaac agtgtatgtt 4381 tacttggaaa agttcatgac ctttcagatg tcactccgaa gagaggatgt gtggcttcca 4441 ctgatgtctt accgaaattc tttgctagct ggtggtgatg atgacaccat gtcagtcatt 4501 agtggaatca gcagccgggg gtcaacagta cggagtaaaa aatcaaaacc atctacagga 4561 aaacggaaag tggttgaggg catgcagctt tcactcactg aagaaagtag tagtagtgac 4621 agtatgtggt taagcagaga acaaacactg cacacccctg ttatgatgca gacaccacaa 4681 ctcacctcca ctattatgag agagcccaaa agattacggc ctgaggatag cttcatgagt 4741 gtttatccaa tgcagactga acatcatcaa acacctcttg attataatcg gcgtggcaca 4801 agcctaatgg aagatgatga agagccaatt gtggaagatg ttatgatgtc ctcagaaggg 4861 aggattgagg atcttaatga gggaatggat tttgacacca tggatataga tttgccacca 4921 tcaaagaaca gacgagagag aacagaactg aagcctgatt tctttgatcc agcttcaatt 4981 atggatgaat cagttcttgg agtgtcaatg tttgattata aagatgacga tgacaagtaa 5041 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat 5101 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa 5161 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact 5221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 5281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac 5341 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc 5401 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 5461 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 5521 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 5581 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 5641 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 5701 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 5761 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga 5821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 5881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg 5941 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 6001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 6061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 6121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 6181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 6241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa 6301 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 6361 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 6421 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 6481 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg 6541 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 6601 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 6661 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 6721 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 6781 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 6841 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 6901 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 6961 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 7021 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 7081 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 7141 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 7201 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 7261 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg 7321 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg 7381 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata 7441 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc 7501 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca 7561 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 7621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 7681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 7741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 7801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 7861 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 7921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 7981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 8041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 8101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 8161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 8221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 8281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 8341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 8401 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca 8461 ttagttcata gccc //