LOCUS Exported 7050 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCAGGS-Raichu-RhoA-CR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7050) AUTHORS Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin MZ TITLE Improving FRET dynamic range with bright green and red fluorescent proteins. JOURNAL Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171. PUBMED 22961245 REFERENCE 2 (bases 1 to 7050) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7050 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..661 /label=chicken beta-actin promoter primer_bind 1594..1616 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind complement(1776..1797) /label=EGFP-N /note="EGFP, reverse primer" CDS 3285..3995 /codon_start=1 /product="monomeric red fluorescent protein derived from mRuby, with improved photophysical properties (Lam et al., 2012)" /label=mRuby2 /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV AKFAGLGGGMDELYK" primer_bind complement(4074..4093) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4139..4194 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4193..4212) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4553..4569) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4553..4569) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4566..4588) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4577..4593 /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(4601..4631) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4646..4667 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4726..4921 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4772..4907 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4834..4853 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4927..5061 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4964..4983) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5018..5037 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5129..5146) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5300..5888) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5380..5399) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6059..6919) /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 6682..6701 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6920..7024) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct 1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc 1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt 1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 1681 ggcaacgtgc tggttgttgt gctgtctcat cattttggca agaattcggc atggtgagca 1741 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa 1801 acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga 1861 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca 1921 ccttcggcta cggcgtggcc tgcttcagcc gctaccccga ccacatgaag cagcacgact 1981 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg 2041 acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca 2101 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt 2161 acaacttcaa cagccacaac gtctatatca cggccgacaa gcagaagaac ggcatcaagg 2221 ctaacttcaa gatccgccac aacgttgagg acggcagcgt gcagctcgcc gaccactacc 2281 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc 2341 atcagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt 2401 tcgtaaccgc cgccctcgag tggtccctgc tagagcagct gggcctggcc ggggcagacc 2461 tggcggcccc cggggtacag cagcagctgg agctggagcg ggagcggctg cggcgggaaa 2521 tccgcaagga gctgaagctg aaggagggtg ctgagaacct gcggcgggcc accactgacc 2581 tgggccgcag cctgggcccc gtagagctgc tgctgcgggg ctcctcgcgc cgcctcgacc 2641 tgctgcacca gcagctgcag gagctgcacg cccacgtgac cggaggtgga accggtggtg 2701 gaggtaccat ggctgccatc cggaagaaac tggtgattgt tggtgatgga gcctgtggaa 2761 agacatgctt gctcatagtc ttcagcaagg accagttccc agaggtgtat gtgcccacag 2821 tgtttgagaa ctatgtggca gatatcgagg tggatggaaa gcaggtagag ttggctttgt 2881 gggacacagc tgggcaggaa gattatgatc gcctgaggcc cctctcctac ccagataccg 2941 atgttatact gatgtgtttt tccatcgaca gccctgatag tttagaaaac atcccagaaa 3001 agtggacccc agaagtcaag catttctgtc ccaacgtgcc catcatcctg gttgggaata 3061 agaaggatct tcggaatgat gagcacacaa ggcgggagct agccaagatg aagcaggagc 3121 cggtgaaacc tgaagaaggc agagatatgg caaacaggat tggcgctttt gggtacatgg 3181 agtgttcagc aaagaccaaa gatggagtga gagaggtttt tgaaatggct acgagagctg 3241 ctctgcaagc tagacgtggg aagaaaaaat ctggtggcgg ccgcatggtg tctaagggcg 3301 aagagctgat caaggaaaat atgcgtatga aggtggtcat ggaaggttcg gtcaacggcc 3361 accaattcaa atgcacaggt gaaggagaag gcaatccgta catgggaact caaaccatga 3421 ggatcaaagt catcgaggga ggacccctgc catttgcctt tgacattctt gccacgtcgt 3481 tcatgtatgg cagccgtact tttatcaagt acccgaaagg cattcctgat ttctttaaac 3541 agtcctttcc tgagggtttt acttgggaaa gagttacgag atacgaagat ggtggagtcg 3601 tcaccgtcat gcaggacacc agccttgagg atggctgtct cgtttaccac gtccaagtca 3661 gaggggtaaa ctttccctcc aatggtcccg tgatgcagaa gaagaccaag ggttgggagc 3721 ctaatacaga gatgatgtat ccagcagatg gtggtctgag gggatacact catatggcac 3781 tgaaagttga tggtggtggc catctgtctt gctctttcgt aacaacttac aggtcaaaaa 3841 agaccgtcgg gaacatcaag atgcccggta tccatgccgt tgatcaccgc ctggaaaggt 3901 tagaggaaag tgacaatgaa atgttcgtag tacaacgcga acacgcagtt gccaagttcg 3961 ccgggcttgg tggtggcatg gacgagctgt acaagtctag aaagatgagc aaagatggta 4021 aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtaagtcgac ggatcttttt 4081 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa 4141 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga 4201 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg 4261 gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta 4321 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta 4381 gatttttttt tatattttgt ttgtgttatt tttttcttta acatccctaa aattttcctt 4441 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc 4501 tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc 4561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 4621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc 4741 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 4801 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 4861 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 4921 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 4981 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 5041 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 5101 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 5161 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 5221 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 5281 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 5341 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 5401 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 5461 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt 5521 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 5581 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 5641 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 5701 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg 5761 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 5821 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 5881 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 5941 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 6001 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 6061 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 6121 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 6181 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 6241 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 6301 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 6361 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6421 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6481 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6541 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 6601 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 6661 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6721 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 6781 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6841 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6901 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 6961 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 7021 cgcgcacatt tccccgaaaa gtgccacctg //