LOCUS Exported 7169 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pmEGFP_a_tubulin_IRES_puro2b SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7169) AUTHORS Steigemann P, Wurzenberger C, Schmitz MH, Held M, Guizetti J, Maar S, Gerlich DW TITLE Aurora B-mediated abscission checkpoint protects against tetraploidization. JOURNAL Cell. 2009 Feb 6. 136(3):473-84. PUBMED 19203582 REFERENCE 2 (bases 1 to 7169) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7169 /organism="synthetic DNA construct" /mol_type="other DNA" intron 74..303 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 286..305 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 361..934 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(528..545) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 755..774 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 967..1566 /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" primer_bind complement(967..986) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1463..1483 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(1702..1719) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1708..1932 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 1979..1998 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(2058..2074) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2058..2074) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2071..2093) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2082..2098 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2106..2136) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2151..2172 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2289..2306) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2460..3048) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2540..2559) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3219..4079) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3842..3861 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4080..4184) /gene="bla" /label=AmpR promoter primer_bind complement(4259..4278) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 4450..4829 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4830..5033 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4984..5004 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 5072..5081 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5078..5794 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK EFVTAAGITLGMDELYK" primer_bind complement(5123..5144) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5384..5403) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" ORIGIN 1 gatccgcggc cgcatagata actgatccag tgtgctggaa ttaattcgct gtctgcgagg 61 gccagctgtt ggggtgagta ctccctctca aaagcgggca tgacttctgc gctaagattg 121 tcagtttcca aaaacgagga ggatttgata ttcacctggc ccgcggtgat gcctttgagg 181 gtggccgcgt ccatctggtc agaaaagaca atctttttgt tgtcaagctt gaggtgtggc 241 aggcttgaga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca 301 caggtgtcca ctcccaggtc caactgcagg tcgagcatgc atctagggcg gccaattccg 361 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg 421 tgcgtttgtc tatatgtgat tttccaccat attgccgtct tttggcaatg tgagggcccg 481 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg 541 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca 601 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct 661 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca 721 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa 781 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg 841 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg 901 ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga 961 ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg 1021 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac 1081 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg 1141 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg 1201 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg 1261 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc 1321 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt 1381 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc 1441 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc 1501 accgccgacg tcgagtgccc gaaggaccgc gcgacctggt gcatgacccg caagcccggt 1561 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc 1621 tccgaccgaa gccgacccgg gcggccccgc cgaccccgca cccgcccccg aggcccaccg 1681 actctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 1741 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 1801 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 1861 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 1921 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgagtgca ttctagttgt 1981 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 2041 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 2101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 2161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 2221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 2281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 2341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 2401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 2461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 2521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 2581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 2641 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 2701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 2761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 2821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 2881 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 2941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 3001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 3061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 3121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 3181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 3241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 3301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 3361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 3421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 3481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 3541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 3601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 3661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 3721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 3781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 3841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 3901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 3961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 4021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 4081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 4141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 4201 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta 4261 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg 4321 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt 4381 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat 4441 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag 4501 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct 4561 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc 4621 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg 4681 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat 4741 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 4801 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 4861 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 4921 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 4981 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag 5041 tgaaccgtca gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct 5101 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt 5161 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat 5221 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg 5281 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc 5341 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa 5401 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg 5461 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag 5521 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat 5581 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc 5641 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct 5701 gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc 5761 cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc gagtgcgtga 5821 gtgcatctcc atccacgttg gccaggctgg tgtccagatt ggcaatgcct gctgggagct 5881 ctactgcctg gaacacggca tccagcccga tggccagatg ccaagtgaca agaccattgg 5941 gggaggagat gactccttca acaccttctt cagtgagacg ggcgctggca agcacgtgcc 6001 ccgggctgtg tttgtagact tggaacccac agtcattgat gaagttcgca ctggcaccta 6061 ccgccagctc ttccaccctg agcagctcat cacaggcaag gaagatgctg ccaataacta 6121 tgcccgaggg cactacacca ttggcaagga gatcattgac cttgtgttgg accgaattcg 6181 caagctggct gaccagtgca ccggtcttca gggcttcttg gttttccaca gctttggtgg 6241 gggaactggt tctgggttca cctccctgct catggaacgt ctctcagttg attatggcaa 6301 gaagtccaag ctggagttct ccatttaccc agcaccccag gtttccacag ctgtagttga 6361 gccctacaac tccatcctca ccacccacac caccctggag cactctgatt gtgccttcat 6421 ggtagacaat gaggccatct atgacatctg tcgtagaaac ctcgatatcg agcgcccaac 6481 ctacactaac cttaaccgcc ttattagcca gattgtgtcc tccatcactg cttccctgag 6541 atttgatgga gccctgaatg ttgacctgac agaattccag accaacctgg tgccctaccc 6601 ccgcatccac ttccctctgg ccacatatgc ccctgtcatc tctgctgaga aagcctacca 6661 tgaacagctt tctgtagcag agatcaccaa tgcttgcttt gagccagcca accagatggt 6721 gaaatgtgac cctcgccatg gtaaatacat ggcttgctgc ctgttgtacc gtggtgacgt 6781 ggttcccaaa gatgtcaatg ctgccattgc caccatcaaa accaagcgca gcatccagtt 6841 tgtggattgg tgccccactg gcttcaaggt tggcatcaac taccagcctc ccactgtggt 6901 gcctggtgga gacctggcca aggtacagag agctgtgtgc atgctgagca acaccacagc 6961 cattgctgag gcctgggctc gcctggacca caagtttgac ctgatgtatg ccaagcgtgc 7021 ctttgttcac tggtacgtgg gtgaggggat ggaggaaggc gagttttcag aggcccgtga 7081 agatatggct gcccttgaga aggattatga ggaggttggt gtggattctg ttgaaggaga 7141 gggtgaggaa gaaggagagg aatactaag //