LOCUS Exported 5198 bp ds-DNA circular SYN 15-JAN-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Untitled SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5198) AUTHORS . TITLE Direct Submission JOURNAL Exported 15 Jan, 2016 from SnapGene Viewer 3.0.1 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5894 /dnas_title="pcDNA5-FRT-TO-V5-SHLD3 for Addgene" /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 235..614 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 820..838 /dnas_title="tetO" /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 841..859 /dnas_title="tetO" /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" polyA_signal 1852..2076 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" protein_bind 2360..2407 /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FRT" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 2415..3437 /dnas_title="HygR" /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /note="HygR" /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGDFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYH WQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSE AMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQS LVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPST RPRAKE" polyA_signal 3567..3688 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(3737..3753) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 3761..3777 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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(3785..3815) /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 3830..3851 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4139..4727) /dnas_title="ori" /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4898..5758) /dnas_title="AmpR" /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" promoter complement(5759..5863) /dnas_title="AmpR promoter" /gene="bla" /note="AmpR promoter" CDS 1059..1805 /note="human SHLD3" /translation="TTEVILHYRPCESDPTQLPKIAEKAIQDFPTRPLSRFIPWFPYD GSKLPLRPKRSPPVISEEAAEDVKQYLTISEHDAKSHSYDCTVDLLEFQPSLKKQHLT WSHTLKEQTNSGNLGKQSEKGKQHKRRSWSISLPSNNCTKNVSPLSKKLQDSLKALNL HSLYRARWTIEHTICNSQTLEDIWTKLNQIIRHNELPSCNATIQRHLGQIWVFCDIMY CEYVGSLLKGRLALTGKINLFVHKYGVIFSM" CDS 990..1805 /note="V5-SHLD3" /translation="MGKPIPNPLLGLDSTAASLNGAPTTEVILHYRPCESDPTQLPKI AEKAIQDFPTRPLSRFIPWFPYDGSKLPLRPKRSPPVISEEAAEDVKQYLTISEHDAK SHSYDCTVDLLEFQPSLKKQHLTWSHTLKEQTNSGNLGKQSEKGKQHKRRSWSISLPS NNCTKNVSPLSKKLQDSLKALNLHSLYRARWTIEHTICNSQTLEDIWTKLNQIIRHNE LPSCNATIQRHLGQIWVFCDIMYCEYVGSLLKGRLALTGKINLFVHKYGVIFSM" CDS 993..1034 /dnas_title="V5 tag" /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttggtacca tgggtaagcc tatccctaac cctctcctcg 1021 gtctcgattc tacggccgcc agtttaaacg gcgcgccaAC TACAGAAGTA ATATTACATT 1081 ATCGACCATG TGAGAGTGAT CCCACACAAC TGCCAAAAAT TGCAGAAAAA GCAATTCAAG 1141 ACTTTCCTAC TCGTCCGCTA TCAAGATTTA TACCTTGGTT TCCATATGAT GGGTCCAAGC 1201 TTCCACTCAG ACCTAAAAGA TCACCACCTG TGATTTCTGA AGAGGCAGCT GAAGATGTGA 1261 AACAGTACTT AACCATTTCA GAACATGATG CTAAGTCACA CAGTTATGAT TGCACAGTAG 1321 ATCTATTGGA GTTTCAACCT AGCTTGAAAA AGCAGCATTT AACCTGGTCA CACACACTGA 1381 AGGAACAGAC TAATTCTGGA AATCTGGGTA AACAATCAGA AAAGGGAAAA CAGCACAAGA 1441 GGAGATCTTG GAGTATTTCC CTTCCCAGCA ATAATTGTAC TAAAAACGTT TCTCCTTTGT 1501 CTAAAAAATT GCAAGATAGT TTAAAGGCAC TAAATTTACA CTCACTTTAT AGAGCAAGAT 1561 GGACAATAGA ACACACTATT TGTAACAGCC AAACTCTGGA AGACATTTGG ACAAAACTCA 1621 ATCAAATTAT TAGGCACAAT GAACTTCCAT CTTGTAATGC TACAATTCAG AGGCATTTAG 1681 GCCAAATATG GGTGTTCTGT GATATTATGT ATTGTGAATA TGTGGGAAGT CTTCTTAAAG 1741 GAAGATTAGC TCTTACTGGA AAAATTAATT TATTTGTGCA TAAATATGGT GTTATTTTTA 1801 GTATGTAGct cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt 1861 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg 1921 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt 1981 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca 2041 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct 2101 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 2161 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 2221 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 2281 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg 2341 gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag 2401 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg 2461 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt 2521 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt 2581 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag 2641 tgcttgacat tggggatttc agcgagagcc tgacctattg catctcccgc cgtgcacagg 2701 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg 2761 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg 2821 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc 2881 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg 2941 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg 3001 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact 3061 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc 3121 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg 3181 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga 3241 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg 3301 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct 3361 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc 3421 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa 3481 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 3541 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat 3601 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 3661 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga 3721 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 3781 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 3841 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 3901 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 3961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 4021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 4081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 4141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 4381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 4441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 4501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 4561 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 4621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 4681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4801 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 4861 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 4921 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4981 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 5041 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 5101 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 5161 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5221 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5281 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5341 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5401 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5461 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5521 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5581 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5641 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5701 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5761 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 5821 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 5881 tgccacctga cgtc //