LOCUS Exported 6136 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses pCAG-Flpo . ACCESSION . VERSION . KEYWORDS pCAG-Flpo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6136) AUTHORS Xue M, Atallah BV, Scanziani M TITLE Equalizing excitation-inhibition ratios across visual cortical neurons. JOURNAL Nature. 2014 Jul 31;511(7511):596-600. doi: 10.1038/nature13321. Epub 2014 Jun 22. PUBMED 25043046 REFERENCE 2 (bases 1 to 6136) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6136 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 5..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 662..1678 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1602..1624 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1686..1705 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1758..1767 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1764..3062 /codon_start=1 /product="nuclear-targeted site-specific recombinase" /label=FLPo /note="FLPo is a mouse codon-optimized fusion between the SV40 nuclear localization signal and a thermostable version of the Saccharomyces cerevisiae site-specific recombinase FLP. Recombination occurs at FRT sequences." /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI" primer_bind complement(3158..3177) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3223..3278 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3277..3296) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3639..3655) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3639..3655) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3652..3674) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3663..3679 /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(3687..3717) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3732..3753 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3812..4007 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3858..3993 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3920..3939 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 4013..4147 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4050..4069) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4104..4123 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4215..4232) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4386..4974) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4466..4485) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5145..6005) /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 5768..5787 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6006..6110) /gene="bla" /label=AmpR promoter ORIGIN 1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat 61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccagggggg gggggggggg gggggggggg 541 gggggggggg gggggggggg gggcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc 721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc 781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga 841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg 901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg 961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg 1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt 1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca 1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg 1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc 1261 ggggccgcct cgggccgggg ggggctcggg ggaggggcgc ggcggccccc ggagcgccgg 1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc 1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac 1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga 1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc 1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt 1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct 1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tctgcagtcg 1741 acggtaccgg atccgccgcc accatggctc ctaagaagaa gaggaaggtg atgagccagt 1801 tcgacatcct gtgcaagacc ccccccaagg tgctggtgcg gcagttcgtg gagagattcg 1861 agaggcccag cggcgagaag atcgccagct gtgccgccga gctgacctac ctgtgctgga 1921 tgatcaccca caacggcacc gccatcaaga gggccacctt catgagctac aacaccatca 1981 tcagcaacag cctgagcttc gacatcgtga acaagagcct gcagttcaag tacaagaccc 2041 agaaggccac catcctggag gccagcctga agaagctgat ccccgcctgg gagttcacca 2101 tcatccctta caacggccag aagcaccaga gcgacatcac cgacatcgtg tccagcctgc 2161 agctgcagtt cgagagcagc gaggaggccg acaagggcaa cagccacagc aagaagatgc 2221 tgaaggccct gctgtccgag ggcgagagca tctgggagat caccgagaag atcctgaaca 2281 gcttcgagta caccagcagg ttcaccaaga ccaagaccct gtaccagttc ctgttcctgg 2341 ccacattcat caactgcggc aggttcagcg acatcaagaa cgtggacccc aagagcttca 2401 agctggtgca gaacaagtac ctgggcgtga tcattcagtg cctggtgacc gagaccaaga 2461 caagcgtgtc caggcacatc tactttttca gcgccagagg caggatcgac cccctggtgt 2521 acctggacga gttcctgagg aacagcgagc ccgtgctgaa gagagtgaac aggaccggca 2581 acagcagcag caacaagcag gagtaccagc tgctgaagga caacctggtg cgcagctaca 2641 acaaggccct gaagaagaac gccccctacc ccatcttcgc tatcaagaac ggccctaaga 2701 gccacatcgg caggcacctg atgaccagct ttctgagcat gaagggcctg accgagctga 2761 caaacgtggt gggcaactgg agcgacaaga gggcctccgc cgtggccagg accacctaca 2821 cccaccagat caccgccatc cccgaccact acttcgccct ggtgtccagg tactacgcct 2881 acgaccccat cagcaaggag atgatcgccc tgaaggacga gaccaacccc atcgaggagt 2941 ggcagcacat cgagcagctg aagggcagcg ccgagggcag catcagatac cccgcctgga 3001 acggcatcat cagccaggag gtgctggact acctgagcag ctacatcaac aggcggatct 3061 gagaattcaa gcttaaccgc actcctcagg tgcaggctgc ctatcagaag gtggtggctg 3121 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt 3181 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt 3241 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg 3301 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata 3361 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc 3421 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat 3481 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag 3541 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga 3601 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga 3661 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 3721 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 3781 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc 3841 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3901 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 3961 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt 4021 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4081 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4141 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 4201 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 4261 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 4321 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 4381 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 4441 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 4501 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 4561 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 4621 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 4681 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 4741 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 4801 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 4861 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 4921 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 4981 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 5041 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 5101 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 5161 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 5221 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 5281 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 5341 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 5401 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 5461 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 5521 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 5581 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 5641 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 5701 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 5761 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 5821 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 5881 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 5941 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 6001 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 6061 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 6121 cgaaaagtgc cacctg //