LOCUS Exported 6922 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses diphtheria toxin A subunit in Cre positive cells, and mCherry in all the infected cells. ACCESSION . VERSION . KEYWORDS pAAV-mCherry-flex-dtA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6922) AUTHORS Wu Z, Autry AE, Bergan JF, Watabe-Uchida M, Dulac CG TITLE Galanin neurons in the medial preoptic area govern parental behaviour. JOURNAL Nature. 2014 May 15;509(7500):325-30. doi: 10.1038/nature13307. PUBMED 24828191 REFERENCE 2 (bases 1 to 6922) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6922 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 92..1270 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 323..1261 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1218..1238 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 1334..1343 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1349..2056 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(1491..1509) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1759..1778 /label=mCherry-F /note="mCherry, forward primer" CDS complement(2170..2742) /codon_start=1 /gene="Corynebacterium diphtheriae tox" /product="diphtheria toxin fragment A" /label=DTA /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI NFETRGKRGQDAMYEYMAQACAGNRVRR" misc_feature 2910..3498 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2963..2983) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3381..3392) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3530..4006 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3729..3748) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4046..4186 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4046..4175 /label=AAV2 ITR rep_origin 4261..4716 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4348..4367) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4558..4579 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4733..4752) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4852..4874 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4912..4930) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4998..5102 /gene="bla" /label=AmpR promoter CDS 5103..5963 /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 complement(5321..5340) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6134..6722 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6623..6642 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 6784..6913 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg 61 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg 121 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta 181 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc 241 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa 301 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt 361 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct 421 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc 481 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat 541 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt 601 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga 661 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca 721 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta 781 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc 841 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc 901 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga 961 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac 1021 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc 1081 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac 1141 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt 1201 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca 1261 ggtgtcgtga ggtaccggat cctctagagt caattcatta cctcgtatag gatactttat 1321 acgaagttat gccgccacca tgggtggggt gagcaagggc gaggaggata acatggccat 1381 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt 1441 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa 1501 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta 1561 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt 1621 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt 1681 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac 1741 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc 1801 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct 1861 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt 1921 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga 1981 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga 2041 cgagctgtac aagtaataac ctcgtatagc atacattata cgaagtaata gatcttagag 2101 ctttaaatct ctgtaggtag tttgtccaat tatgtcacac cacagaagta aggttccttc 2161 acaaagagat cgcctgacac gatttcctgc acaggcttga gccatatact catacatcgc 2221 atcttggcca cgttttccac gggtttcaaa attaatctca agttctacgc ttaacgcttt 2281 cgcctgttcc cagttattaa tatattcaac gctagaactc ccctcagcga agggaaggct 2341 gagcactaca cgcgaagcac catcaccgaa ccttttgata aactcttccg ttccgacttg 2401 ctccatcaac ggttcagtga gacttaaacc taactctttc ttaatagttt cggcattatc 2461 cacttttagt gcgagaacct tcgtcagtcc tggatacgtc actttgacca cgcctccagc 2521 ttttccagag agcgggtttt cattatctac agagtatccc gcagcgtcgt atttattgtc 2581 ggtactataa aaccctttcc aatcatcgtc ataatttcct tgtgtaccag attttggctt 2641 ttgtatacct ttttgaatgg aatctacata accaggttta gtcccgtggt acgaagaaaa 2701 gttttccatc acaaaagatt tagaagaatc aacaacatca tcaggatcca tggcgaggac 2761 gtcgacataa cttcgtataa agtatcctat acgaggtaat ttgccttaac ccagaaatta 2821 tcagtactat tctttagaat ggtgcaaaga attacttcgt ataatgtatg ctatacgagg 2881 ttatcgcatg ctagatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa 2941 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 3001 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 3061 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 3121 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 3181 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 3241 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 3301 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga 3361 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 3421 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 3481 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc 3541 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 3601 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 3661 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 3721 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 3781 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 3841 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 3901 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 3961 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt 4021 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 4081 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 4141 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 4201 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 4261 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 4321 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 4381 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 4441 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 4501 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 4561 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat 4621 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 4681 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 4741 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 4801 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 4861 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 4921 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 4981 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 5041 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 5101 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 5161 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 5221 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 5281 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 5341 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 5401 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 5461 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 5521 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 5581 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 5641 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 5701 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 5761 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 5821 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 5881 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 5941 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 6001 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 6061 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 6121 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 6181 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 6241 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 6301 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 6361 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 6421 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 6481 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 6541 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 6601 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 6661 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 6721 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 6781 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc 6841 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc 6901 actaggggtt cctgcggccg ca //