LOCUS Exported 6695 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre-dependent expression of FlpO from CAV2 virus. ACCESSION . VERSION . KEYWORDS pTCAV-FLEx(loxP)-FlpO SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6695) AUTHORS Schwarz LA, Miyamichi K, Gao XJ, Beier KT, Weissbourd B, DeLoach KE, Ren J, Ibanes S, Malenka RC, Kremer EJ, Luo L TITLE Viral-genetic tracing of the input-output organization of a central noradrenaline circuit. JOURNAL Nature. 2015 Aug 6;524(7563):88-92. doi: 10.1038/nature14600. Epub 2015 Jul 1. PUBMED 26131933 REFERENCE 2 (bases 1 to 6695) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6695 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(304..323) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1275..2069) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1386..1405) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 1996..2015 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(2118..2495) /label=ISS /note="immunostimulatory sequence from the AmpR gene; contains unmethylated CpG dinucleotides in the context of 5'-AACGTT-3' (Sato et al., 1996)" primer_bind 2442..2461 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2680..2784) /gene="bla" /label=AmpR promoter primer_bind 2852..2870 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" enhancer 3621..3906 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 3907..4118 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4069..4089 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4115..4139 /label=LNCX /note="Human CMV promoter, forward primer" intron 4279..4411 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 4394..4413 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 4456..4474 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind complement(4512..4545) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(4608..4641) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 5935..5968 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 6039..6072 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter complement(6114..6131) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 6149..6270 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6160..6179) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6214..6233 /label=SV40pA-R /note="SV40 polyA, reverse primer" ORIGIN 1 cggccttaat taaggcgcgc ccgggcattt aaatgtttaa acggccgcag atctgcgacg 61 cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc 121 gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaaggc 181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 301 tataaagata ccaggcgttt cccctggaaa gctcccttgt gcgctctcct gttccgaccc 361 tgccgttacc ggatacctgt cccgctttct cctttgggaa gcgtggcgct ttcttatagc 421 tcacgcttgt aggtatctta tcggtgtagt cgttcgctca agctgggctg tgtgcacgaa 481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 901 ttcacctaga tccttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcttcgaa 1261 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa 1321 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct 1381 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg 1441 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca 1501 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac 1561 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg 1621 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag 1681 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg 1741 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag 1801 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc 1861 agccacgata gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc 1921 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag 1981 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa 2041 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg 2101 atctttgcaa aagccgcaac gttgttgcca ttgtgcaggc atcgtggtgt cacgctcgtc 2161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 2221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 2281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 2341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 2401 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag 2461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 2521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 2581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 2641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 2701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 2761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga 2821 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct 2881 tcaagaattg gatccgaatt gatagaacta ttacagacag acaacgctgg acacggtatt 2941 atcattcaaa aaagaaataa attttatttt tacattaatc tcgggcgatt cgttgcttta 3001 gatcatttac ttctcgctga atagcagtca cagagttttg catggcagcc acctggatct 3061 tcagctcatc caaaagaaca ttcaccgcaa gcatgttcag ctgctgctcc tcatacaaag 3121 ttctgatgcg cggggctgca agcgggcgtc cactttccag cacatctgag ggcacgggct 3181 ttccttctaa atctgacccg gtgacattct gtcttgctcc tgcccaagac ggaatacgcg 3241 tagtcacaaa acacgtgttc acaagcccct tctgttgagg gtccatggct gtaaatacac 3301 agccacgtac cgcgcccctt ttatactccc ccaaaccacg cccatctact cacctaaaaa 3361 ttatcttcat cactgctaaa gaactcagcc gtgtcttgag agttcagctc ccggtcaatc 3421 ctggtgtcag ttacataggt ggcccgcagg gaaggggtgc tgtgtctgaa cccgcaagta 3481 cacagtcttt cactagcagc attgcctgcc tcgcccagtt gcataatggc acaagtagca 3541 tcatacacac caaacaaata aattctctgc acaacccccc gaggggctgc aattacacac 3601 attgacaaag taggggtact cgttacataa cttacggtaa atggcccgcc tggctgaccg 3661 cccaacgacc cccgcccatt gacgtcaata gtaacgccaa tagggacttt ccattgacgt 3721 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 3781 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 3841 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 3901 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg 3961 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 4021 cgggactttc caaaatgtcg taacaactgc gatcgcccgc cccgttgacg caaatgggcg 4081 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatca 4141 ctagaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg 4201 acacaacagt ctcgaactta agctgcagtg actctcttaa ggtagccttg cagaagttgg 4261 tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag 4321 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt 4381 actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct 4441 taaggctaga gtacttaata cgactcacta taggcctagc ctcgagaatt cgcagtcgac 4501 ggtaccccgg gataacttcg tataaagtat cctatacgaa gttatatcaa aataggaaga 4561 ccaatgcttc accatcgacc cgaattgcca agcatcacca tcgacacata acttcgtata 4621 atgtatgcta tacgaagtta tgtcgactca tcagatccgc ctgttgatgt agctgctcag 4681 gtagtccagc acctcctggc tgatgatgcc gttccaggcg gggtatctga tgctgccctc 4741 ggcgctgccc ttcagctgct cgatgtgctg ccactcctcg atggggttgg tctcgtcctt 4801 cagggcgatc atctccttgc tgatggggtc gtaggcgtag tacctggaca ccagggcgaa 4861 gtagtggtcg gggatggcgg tgatctggtg ggtgtaggtg gtcctggcca cggcggaggc 4921 cctcttgtcg ctccagttgc ccaccacgtt tgtcagctcg gtcaggccct tcatgctcag 4981 aaagctggtc atcaggtgcc tgccgatgtg gctcttaggg ccgttcttga tagcgaagat 5041 ggggtagggg gcgttcttct tcagggcctt gttgtagctg cgcaccaggt tgtccttcag 5101 cagctggtac tcctgcttgt tgctgctgct gttgccggtc ctgttcactc tcttcagcac 5161 gggctcgctg ttcctcagga actcgtccag gtacaccagg gggtcgatcc tgcctctggc 5221 gctgaaaaag tagatgtgcc tggacacgct tgtcttggtc tcggtcacca ggcactgaat 5281 gatcacgccc aggtacttgt tctgcaccag cttgaagctc ttggggtcca cgttcttgat 5341 gtcgctgaac ctgccgcagt tgatgaatgt ggccaggaac aggaactggt acagggtctt 5401 ggtcttggtg aacctgctgg tgtactcgaa gctgttcagg atcttctcgg tgatctccca 5461 gatgctctcg ccctcggaca gcagggcctt cagcatcttc ttgctgtggc tgttgccctt 5521 gtcggcctcc tcgctgctct cgaactgcag ctgcaggctg gacacgatgt cggtgatgtc 5581 gctctggtgc ttctggccgt tgtaagggat gatggtgaac tcccaggcgg ggatcagctt 5641 cttcaggctg gcctccagga tggtggcctt ctgggtcttg tacttgaact gcaggctctt 5701 gttcacgatg tcgaagctca ggctgttgct gatgatggtg ttgtagctca tgaaggtggc 5761 cctcttgatg gcggtgccgt tgtgggtgat catccagcac aggtaggtca gctcggcggc 5821 acagctggcg atcttctcgc cgctgggcct ctcgaatctc tccacgaact gccgcaccag 5881 caccttgggg ggggtcttgc acaggatgtc gaactggctc atggtggcgc gcctataact 5941 tcgtatagga tactttatac gaagttatca ttgggattct tcctattttg atccaagcat 6001 caccatcgac cctctagtcc agatctcacc atcgacccat aacttcgtat agcatacatt 6061 atacgaagtt ataagcttaa ttactagatc atgcatgctg tacaggccgc ttccctttag 6121 tgagggttaa tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca 6181 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt 6241 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt 6301 caggttcagg gggaggtgtg ggaggttttt tagtcgacta ggtggggtgc tgtaaacaag 6361 aacagagtgt ttctgttata ttgacataaa ctttgacatt agccgtggag gaactggcac 6421 caaaaccagc tccataaaca actaaatctt ccgcaaacag tggtaaaagc accacccgct 6481 aaaccgcaaa atcaataaac gcccaaaaac ccggggcgcc ggccaaaagt ccgcggaact 6541 cgccctgtcg taaaaccacg cctttgacgt cactggacat tcccgtggga acaccctgac 6601 cagggcgtga cctgaacctg accgtcccat gaccccgccc cttgcaacac ccaaatttaa 6661 gccacacctc tttgtcctgt atattattga tgatg //