LOCUS Exported 7274 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS PSD-95 FLAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7274) AUTHORS Zhang J, Vinuela A, Neely MH, Hallett PJ, Grant SG, Miller GM, Isacson O, Caron MG, Yao WD TITLE Inhibition of the dopamine D1 receptor signaling by PSD-95. JOURNAL J Biol Chem. 2007 May 25. 282(21):15778-89. PUBMED 17369255 REFERENCE 2 (bases 1 to 7274) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7274 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 110..489 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 490..693 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 644..664 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 695..713 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 716..734 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 744..768 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 865..874 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3049..3072 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(3101..3118) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3107..3331 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3464..3483) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3615..3662 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 3670..4692 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4822..4943 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4859..4878) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4913..4932 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4992..5008) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4992..5008) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5005..5027) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5016..5032 /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(5040..5070) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5085..5106 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5223..5240) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5394..5982) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5474..5493) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6153..7013) /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 6776..6795 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7014..7118) /gene="bla" /label=AmpR promoter primer_bind complement(7193..7212) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg 61 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 121 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 181 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 241 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 301 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 361 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 421 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 481 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 541 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 601 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 661 cgtgtacggt gggaggtcta tataagcaga gctctcccta tcagtgatag agatctccct 721 atcagtgata gagatcgtcg acgagctcgt ttagtgaacc gtcagatcgc ctggagacgc 781 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccggactcta 841 gcgtttaaac ttaagcttgg taccgccacc atggactgtc tctgtatagt gacaaccaag 901 aaataccgct accaagatga agacacgccc cctctggaac acagcccggc ccacctcccc 961 aaccaggcca attctccccc tgtgattgtc aacacggaca ccctagaagc cccaggatat 1021 gagttgcagg tgaatggaac agagggggag atggagtatg aggagatcac attggaaagg 1081 ggtaactcag gtctgggctt cagcatcgca ggtggcgctg acaacccgca catcggtgac 1141 gacccgtcca tttttatcac caagatcatt cctggtgggg ctgcagccca ggatggccgc 1201 ctcagggtca atgacagcat cctgtttgta aatgaagtgg atgttcggga ggtgacccat 1261 tcagctgcgg tggaggccct caaagaggca ggttccatcg ttcgcctcta tgtcatgcgc 1321 cggaaacccc cagccgaaaa ggtcatggag atcaaactca tcaaagggcc taaaggactt 1381 ggcttcagca ttgcgggggg cgttgggaac cagcacatcc ctggagataa cagcatctat 1441 gtaacgaaga tcatcgaagg aggtgctgcc cacaaggatg gcaggttgca gattggagac 1501 aagatcctgg cggtcaacag tgtggggctg gaggacgtca tgcacgagga tgccgtggca 1561 gccctgaaga acacatatga cgttgtgtac ctaaaggtgg ccaagcccag caatgcctac 1621 ctgagtgaca gctatgctcc cccagacatc acaacctcgt attctcagca cctggacaat 1681 gagatcagtc atagcagcta cttgggcact gactacccca cagccatgac ccccacttcc 1741 cctcggcgct actcccctgt ggccaaggac ctgctggggg aggaagacat tccccgggaa 1801 ccaaggcgga tcgtgatcca tcggggctcc accggcctgg gcttcaacat cgtgggcggc 1861 gaggatggtg aaggcatctt catctccttc atccttgctg ggggtccagc cgacctcagt 1921 ggggagctac ggaaggggga ccagatcctg tcggtcaatg gtgttgacct ccgcaatgcc 1981 agtcacgaac aggctgccat tgccctgaag aatgcgggtc agacggtcac gatcatcgct 2041 cagtataaac cagaagagta tagtcgattc gaggccaaga tccatgatct tcgggaacag 2101 ctcatgaata gtagcctagg ctcagggact gcatccttgc gaagcaaccc caagaggggc 2161 ttctacatta gggccctgtt tgattacgac aagaccaagg actgcggttt cttgagccag 2221 gccctgagct tccgcttcgg ggatgtgctt catgtcattg acgctggtga cgaagagtgg 2281 tggcaagcac ggcgggtcca ctccgacagt gagaccgacg acattggctt cattcccagc 2341 aaacggcggg tcgagcgacg agagtggtca aggttaaagg ccaaggactg gggctccagc 2401 tctggatcac agggtcgaga agactcggtt ctgagctatg agacggtgac ccagatggaa 2461 gtgcactatg ctcgtcccat catcatcctt ggacccacca aagaccgtgc caacgatgat 2521 cttctctccg agttccccga caagtttgga tcctgtgtcc ctcatacgac acgtcctaag 2581 cgggaatatg agatagacgg ccgggattac cactttgtct cctcccggga gaaaatggag 2641 aaggacatcc aggcacacaa gttcattgag gctggccagt acaacagcca cctctatggg 2701 accagcgtcc agtctgtgcg agaggtagca gagcagggga agcactgcat cctcgatgtc 2761 tcggccaatg ccgtgcggcg gctgcaggcg gcccacctgc accccatcgc catcttcatc 2821 cgtccccgct ccctggagaa tgtgctagag atcaataagc ggatcacaga ggagcaagcc 2881 cggaaagcct tcgacagagc cacgaagctg gagcaggagt tcacagagtg cttctcagcc 2941 atcgtagagg gcgacagctt tgaagagatc tatcacaaag tgaaacgtgt cattgaagac 3001 ctctcaggcc cctacatctg ggtcccagcc cgagagagac tcctcgagga ctacaaggac 3061 gacgatgaca agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt 3121 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 3181 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 3241 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 3301 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 3361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 3541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3601 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact 3661 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag 3721 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc 3781 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac 3841 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt 3901 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc 3961 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc 4021 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg 4081 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat 4141 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc 4201 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat 4261 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc 4321 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg 4381 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga 4441 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg 4501 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga 4561 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc 4621 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg 4681 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg 4741 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4801 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 4861 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4921 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 4981 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5041 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5101 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 5161 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 5221 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 5281 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 5341 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5401 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 5461 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 5521 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 5581 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 5641 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 5701 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 5761 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 5821 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 5881 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 5941 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6001 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6061 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6121 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6181 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6241 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6301 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6361 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6421 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 6481 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 6541 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6601 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 6661 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 6721 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 6781 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 6841 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6901 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 6961 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7021 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7081 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7141 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 7201 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 7261 agtagtgcgc gagc //