LOCUS Exported 6601 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FLX1.8MitoKate2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6601) TITLE Owens lab FIV plasmids REFERENCE 2 (bases 1 to 6601) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6601 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 169..191 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 183..200 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 184..200 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 266..569 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 570..773 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 724..744 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 1597..1900 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" CDS 2105..2179 /codon_start=1 /product="mitochondrial presequence of human cytochrome c oxidase subunit VIII (Rizutto et al., 1989)" /label=COX8 presequence /translation="MSVLTPLLLRGLTGSARRLPVPRAK" regulatory 2207..2216 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2213..2911 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" misc_feature 2918..3506 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2971..2991) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3389..3400) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3744..3760) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3744..3760) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(4174..4190) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4174..4190) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4187..4209) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4198..4214 /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(4222..4252) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4267..4288 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4349..4937) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4429..4448) /label=pBR322ori-F /note="pBR322 origin, forward primer" rep_origin 5048..5557 /direction=RIGHT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5135..5154) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5341..5362 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(5599..6459) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6222..6241 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6460..6564) /gene="bla" /label=AmpR promoter ORIGIN 1 aaaaaaaagc ccgccgaagc gggctttatt accaagcgaa gcgccattcg ccattcaggc 61 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga 121 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac 181 gttgtaaaac gacggccagt ccgtaatacg actcacttaa ggccttgact agagggtacc 241 agagctcacc taggtatcta gaccgcgtta cataacttac ggtaaatggc ccgcctggct 301 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc 361 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg 421 cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat 481 ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca 541 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc 601 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 661 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt 721 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctgtgaa 781 acttcgagga gtctctttgt tgaggacttt tgagttctcc cttgaggctc ccacagatac 841 aataaatatt tgagattgaa ccctgtcgag tatctgtgta atctttttta cctgtgaggt 901 ctcggaatcc gggccgagaa cttcgcagtt ggcgcccgaa cagggacttg attgagagtg 961 attgaggaag tgaagctaga gcaatagaaa gctgttaagc agaactcctg ctgacctaaa 1021 tagggaagca gtagcagacg ctgctaacag tgagtatctc tagtgaagcg gactcgagct 1081 cataatcaag tcattgttta aaggcccaga taaattacat ctggtgactc ttcgcggacc 1141 ttcaagccag gagattcgcc gagggacagt caacaaggaa ggagagattc tacagcaact 1201 tggggattgg acaggggcga gattggaaaa tggccattaa gagatgtagt aatgttgctg 1261 taggagtagg ggggaagagt aaaaaatttg gagaagggaa tttcagatgg gccattagaa 1321 tggctaatgt atctacagga cgagaacctg gtgatatacc agagacttta gatcaactaa 1381 ggttggttat ttgcgattta caagaaagaa gagaaaaatt tggatctggc aaagaaattg 1441 atatggcaat tgtgacatta aaagtctttg cggtagcagg acttttaaat atgacggtgt 1501 cttactagcc aattttaaaa gaagaggtag gataggaggg gaattcttaa gaatacaaga 1561 ttatttttct gcaataccac aaaaattgat gcatcgcgtt acataactta cggtaaatgg 1621 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 1681 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 1741 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 1801 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 1861 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta 1921 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga 1981 cgtcaatggg agtttgtttt gagatctgaa ggggggctat aaaagcgatg gatccgagct 2041 cggccctcat tctggagact ctagaggcct tgaattccgc ggatccgcta gcgctacgcg 2101 taccatgtcc gtcctgacgc cgctgctgct gcggggcttg acaggctcgg cccggcggct 2161 cccagtgccg cgcgccaaga tccattcgtt gggggatcca ccggtcgcca ccatggtgag 2221 cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca 2281 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag 2341 aatcaaggcg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt 2401 catgtacggc agcaaaacct tcatcaacca cacccagggc atccccgact tctttaagca 2461 gtccttcccc gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct 2521 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag 2581 aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc 2641 ctccaccgag accctgtacc ccgctgacgg cggcctggaa ggcagagccg acatggccct 2701 gaagctcgtg ggcgggggcc acctgatctg caacttgaag accacataca gatccaagaa 2761 acccgctaag aacctcaaga tgcccggcgt ctactatgtg gacagaagac tggaaagaat 2821 caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg 2881 cgacctccct agcaaactgg ggcacagatg agtcgacaat caacctctgg attacaaaat 2941 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc 3001 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt 3061 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg 3121 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg 3181 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc 3241 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt 3301 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct 3361 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg 3421 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg 3481 gatctccctt tgggccgcct ccccgcctgc tcgacgaatc aaatcaaact aataaagtat 3541 gtattgtaag gtaaaaggaa aagacaaaga agaagaagaa agaagaaagc cttcaagagg 3601 atgatgacag agttagaaga tcgcttcagg aagctatttg gcacgacttc tacaacggga 3661 gacagcacag tagattctga agatgaacct cctaaaaaag aaaaaagggt ggactgggat 3721 gagtattgga atgcagcccg ggggatccac tagttctaga gcggccgcag gagtctcttt 3781 gttgaggact tttgagttct cccttgaggc tcccacagat acaataaata tttgagattg 3841 aaccctgtcg agtatctgtg taatcttttt tacctgtgag gtctcggaat ccgggccgag 3901 aacttcgcag gagagattac agaatggaag gaaaggacca gcacagttca cagcacttca 3961 agtgcctgga agtttaaaaa aggggtggtg tggtgtatgt aagtgtaact actatccctt 4021 ccccaattct gttcagggcc atcatgtcag cacccacctc tgatccagag aagtcaaggt 4081 tcagggggac taaaaagcag ccctcctcac ggtagtaaca gtatggatct gcccactagt 4141 gggagtcgta ttacgtagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 4201 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 4261 tgcctaatga gtgagctaac tcacattaca tgtgagcaaa aggccagcaa aaggccagga 4321 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 4381 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 4441 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 4501 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 4561 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 4621 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 4681 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 4741 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 4801 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 4861 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 4921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 4981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 5041 tccttttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 5101 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 5161 ctcgccacgt tcgctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 5221 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg 5281 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 5341 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt 5401 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 5461 ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aatatttgct tatacaatct 5521 tcctgttttt ggggcttttc tgattatcaa ccggggtaaa tcaatctaaa gtatatatga 5581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6181 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag 6241 cagaacttta aaagtgctca tcattggaga acgttcttcg gggcgaaaac tctcaaggat 6301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6361 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6421 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtagttaa 6601 c //