LOCUS Exported 5695 bp ds-DNA circular SYN 05-SEP-2019 DEFINITION pcDNA3.1ZEO.seq, 5015 bases, 0 checksum. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5695) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, Sep 5, 2019 from SnapGene 4.3.11 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5695 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 943..961 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 981..990 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 987..1049 /codon_start=1 /label=GFP10 /translation="MGDLPDDHYLSTQTILSKDLN" CDS 1056..1085 /codon_start=1 /label=linker /translation="GGGGSGGGGS" CDS 1092..1670 /codon_start=1 /product="human Rho family small GTPase RhoA" /label=RhoA /translation="AAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADI EVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSL?NIP?KWTPEVKH FCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTK DGVREVFEMATRAALQARRGKKKSGCLVL" polyA_signal 1708..1932 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1978..2406 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2420..2750 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2601..2736 /label=SV40 ori /note="SV40 origin of replication" promoter 2798..2845 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 2864..3238 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 3368..3489 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3538..3554) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 3562..3578 /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(3586..3616) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3631..3652 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3940..4528) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4699..5559) /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" promoter complement(5560..5664) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag 961 ggagacccaa gctggctagc gccaccatgg gcgacctgcc cgacgaccac tacctgtcca 1021 cccagaccat cctgagcaag gacctgaaca tcgatggtgg cggtggctct ggaggtggtg 1081 ggtcctccgg agctgccatc cgcaagaaac tggtgattgt tggtgatgga gcctgtggaa 1141 agacatgctt gctcatagtc ttcagcaagg accagttccc agaggtgtat gtgcccacag 1201 tgtttgagaa ctatgtggca gatatcgagg tggatggaaa gcaggtagag ttggctttgt 1261 gggacacagc tgggcaggaa gattatgatc gcctgaggcc cctctcctac ccagataccg 1321 atgttatact gatgtgtttt tccatcgaca gccctgatag tttanaaaac atcccanaaa 1381 agtggacccc agaagtcaag catttctgtc ccaacgtgcc catcatcctg gttgggaata 1441 agaaggatct tcggaatgat gagcacacaa ggcgggagct agccaagatg aagcaggagc 1501 cggtgaaacc tgaagaaggc agagatatgg caaacaggat tggcgctttt gggtacatgg 1561 agtgttcagc aaagaccaaa gatggagtga gagaggtttt tgaaatggct acgagagctg 1621 ctctgcaagc tagacgtggg aagaaaaaat ctggttgcct tgtcttgtaa tctagagggc 1681 ccgtttaaac ccgctgatca 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 ctctaggggg tatccccacg 1981 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 2041 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 2101 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg 2161 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 2221 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 2281 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 2341 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 2401 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg 2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 2521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 2581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 2641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 2701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 2761 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata 2821 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag 2881 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg 2941 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt 3001 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt 3061 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt 3121 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt 3181 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca 3241 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt 3301 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc 3361 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 3421 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 3481 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc 3541 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 3601 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 3661 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 3721 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 3781 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 3841 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 3901 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 3961 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4021 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4081 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 4141 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 4201 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 4261 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 4321 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 4381 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 4441 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 4501 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 4561 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 4621 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 4681 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 4741 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 4801 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 4861 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 4921 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 4981 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5041 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 5101 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 5161 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 5221 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 5281 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 5341 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 5401 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 5461 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 5521 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 5581 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 5641 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //