LOCUS Exported 7746 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Encodes the G protein to enable transsynaptic RAV infection. ACCESSION . VERSION . KEYWORDS DIO-EF1a-FLEX-H2B-3XHA-P2A-N2c(G) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7746) AUTHORS Reardon TR, Murray AJ, Turi GF, Wirblich C, Croce KR, Schnell MJ, Jessell TM, Losonczy A TITLE Rabies Virus CVS-N2c Strain Enhances Retrograde Synaptic Transfer and Neuronal Viability. JOURNAL Neuron. 2016 Jan 19. pii: S0896-6273(16)00005-2. doi: 10.1016/j.neuron.2016.01.004. PUBMED 26804990 REFERENCE 2 (bases 1 to 7746) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7746 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR promoter 242..1419 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 473..1410 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1367..1387 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1450..1483 /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 1534..1567 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(3164..3220) /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS complement(3230..3319) /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA" CDS complement(3338..3715) /codon_start=1 /gene="HIST1H2BJ" /product="human histone H2B" /label=H2B /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP GELAKHAVSEGTKAITKYTSSK" protein_bind complement(3731..3764) /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(3815..3848) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3873..4461 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3926..3946) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4344..4355) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4493..4969 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4692..4711) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 5009..5149 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5009..5138 /label=AAV2 ITR rep_origin 5224..5679 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5311..5330) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5521..5542 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5696..5715) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5815..5837 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5875..5893) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5961..6065 /gene="bla" /label=AmpR promoter CDS 6066..6926 /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(6284..6303) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7097..7685 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7586..7605 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt 181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat 241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg 301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag 361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc 421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc 481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat 541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc 1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga 1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg 1561 aagttatggc gcgccttact tgtacatctt acagtctgat ctcacctcca ctcttatatg 1621 attcccatga aggtatgact tttccgcttt gggaagtgac tgacacattc ccccctgtcc 1681 ctccaaaact gcgttgtttc gattctggtc gattggctct tctgcaccat gtcattaggg 1741 aaaatatcaa caccaggcca atcatggccc ctgcagtcat caatacatac tttccccagt 1801 tcgggagacc caggtcaacc cctgagatct gtttgtacac atcggggagg tgaacttcaa 1861 caaaatcctc agcctcatca ccttctttga aaactgtaga agggtctgcc agggggtgca 1921 tcagggggat aactgaagat tccaacaact ccatatgttg ctggaggagg gatgattgca 1981 tctctgggat taggacatgg tcgtcaggcc ctaatattat accattgaaa aacaccccgt 2041 tcacatgagg atggcacctt cctccaactt tcaaacaccc ttttgagggg atgatctcat 2101 tccaggtccg gactgacttg tagtgagcat cagcctccat caaggttttg ttgaatatgg 2161 tatatgcttt tccaaaccct gggacaagtt ttctcaggtg actgagacgt ctgaaactta 2221 ctgacttggt ggtcatgatg gactctaatg cgtccagaca ttcctctctt ttcttgacta 2281 actcctccac aacgagatgc tcaatctcgt ctgagcgaaa gtcgtgcaaa ttcaccaact 2341 gatctggagg gcaccatttg gtctcatctg atgtttgcat cgcgacccat gttccatcca 2401 taagtctaag tccaagaact ccacataact tgagcctgca tgctcctttt agagacttat 2461 acaggcctct ttcatccaca aagccgcaag tcttgttccc gttggatgct ctcttccctc 2521 tgctattggt aaaaatgtca caaggtgtcc ttggtctcgg attctcgggc atccaaatgg 2581 tgtaatcatg gttagttgag cagtaggtag aggacaccgt tattcctgag cactttccgc 2641 cagggaagac ccttgagtga agggatttgt catatgggtc caaatctgtc acacttgggg 2701 atatgataat gagggactct ttggtggttc ttacagttcg aagccagtgg tagtcggggt 2761 atggattgtg tagggactct tcatatctgg ggtcaccggc catcttccag ttatacgcgg 2821 ctctacatgc gtctggggtg gggcggaaat gctttctctt gaatgtggtt gtgacataac 2881 caacaaagtt ggtgtaggtc tctgcctctg tcacaacacc tgtgcaagtg aacccgttca 2941 ctttgatggc tgagatgtat cccactttga gttccatgta ggagaactcg gacaggttgg 3001 tacatccttc atcctccaca accaggttat ttggacagct gagatggtgt atgtcaatag 3061 ggctccaggg accaagttcg tctggtatcg tgtaaatggg gaacttcccg aaacacaacg 3121 aaaaacccag aaggagtaca aacaaaagaa cctgaggaac cataggtcca gggttctcct 3181 ccacgtctcc agcctgcttc agcaggctga agttagtagc tccgcttcca gcgtaatctg 3241 gaacgtcata tggataggat cctgcatagt ccgggacgtc atagggatag cccgcatagt 3301 caggaacatc gtatgggtaa acggcgccgg atccatcctt agagctggtg tacttggtga 3361 tggccttagt accctcggac acggcgtgct tggccaactc cccaggcagc agcaggcgca 3421 cggccgtctg gatctccctg gaggtgatgg tcgagcgctt gttgtaatgc gccaggcggg 3481 aagcctcacc tgcgatgcgc tcgaaaatgt cgttcacaaa cgagttcatg atgcccatgg 3541 ccttggacga aatgccggtg tcagggtgga cctgcttcag aaccttgtac acatagatgg 3601 aatagctctc cttgcggctg cgcttgcgct tcttgccgcc tttcttctgc gccttagtca 3661 ccgccttctt ggagcccttt ttcggggcgg gagcagactt cgctggctct ggcatggtgg 3721 cggctctagc ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa 3781 attatcactg ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac 3841 gaagttatga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg 3901 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 3961 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 4021 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 4081 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 4141 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 4201 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 4261 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg 4321 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 4381 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 4441 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca 4501 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac 4561 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa 4621 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag 4681 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc 4741 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt 4801 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac 4861 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc 4921 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta 4981 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc 5041 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc 5101 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg 5161 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat 5221 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 5281 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 5341 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 5401 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg 5461 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 5521 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt 5581 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 5641 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa 5701 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc 5761 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga 5821 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg 5881 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg 5941 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 6001 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 6061 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 6121 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 6181 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 6241 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 6301 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 6361 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 6421 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 6481 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 6541 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 6601 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 6661 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 6721 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 6781 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 6841 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 6901 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 6961 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 7021 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 7081 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 7141 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 7201 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 7261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 7321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 7381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 7441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 7501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 7561 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 7621 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 7681 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 7741 acatgt //