LOCUS Exported 6643 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV vector that expresses double floxed cyan pre-eGRASP(p30) under the Ef1a promoter.. ACCESSION . VERSION . KEYWORDS pAAV-EWB-DIO-cyan pre-eGRASP(p30) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6643) AUTHORS Choi JH, Sim SE, Kim JI, Choi DI, Oh J, Ye S, Lee J, Kim T, Ko HG, Lim CS, Kaang BK TITLE Interregional synaptic maps among engram cells underlie memory formation. JOURNAL Science. 2018 Apr 27;360(6387):430-435. doi: 10.1126/science.aas9204. PUBMED 29700265 REFERENCE 2 (bases 1 to 6643) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6643 /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)." primer_bind 2787..2808 /label=EGFP-N /note="EGFP, reverse primer" regulatory 2922..2931 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2936..2969) /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(3020..3053) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3073..3661 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3126..3146) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3544..3555) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3674..3898 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 3906..4046 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3906..4035 /label=AAV2 ITR rep_origin 4121..4576 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4208..4227) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4418..4439 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4593..4612) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4712..4734 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4772..4790) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4858..4962 /gene="bla" /label=AmpR promoter CDS 4963..5823 /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(5181..5200) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5994..6582 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6483..6502 /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 gcgccttaga cataatactc cttatccttg ttcttcttgt ttttgttggc 1621 gcttttcgca ctgctgggct gcttctcctt gaccacagcc ccattggact gtgctgagtt 1681 actgatgtag tttcgactct catccacgtg atatgagcct tcatcccggt ttctgtactt 1741 gtacatggca tagaggagga tgaggatgca cagagctgct gctgcgacaa tccccaccac 1801 catgccagtg gtactgctag actcccggat cacctctgcc gagcctgggt atggttcgcg 1861 gccgcccact cgggtggggt tggctaaccc acctgagctc ggctcacagg ggtcaatgtc 1921 ctcatcgtcg ctgggacact ctgccgaggc cacaaggata tcatccgtgg tctggctgat 1981 aggctccttt gtggggggct ttcctcgtcg agctgtgctg gtagccaggg ttgtggtggt 2041 ctccatgatt gaggtggaca tctcggactg catggcagtg gctgtcgact cagttgtcat 2101 agaggaaggc acttcggtac cgcttccgcc tccggaacct cctccaggtg gcagtggtgg 2161 aggctttgta ggagctctag atccgccact gcctccagat ccaccggtct tctcgttggg 2221 gtctttgctc agcacgctct gggtgctcag gtagtggttg tcgggcagca gcacggggcc 2281 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctccac 2341 gttgtggcgc acggtgaagt tggccttgat gccgttcttc tgcttgtcgg cggtgatata 2401 gacgttgccg ctgttgaagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt 2461 gaagtcggtg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcaccac 2521 ggcgcgggtc ttgtacttgc cgtcgtcctt gaaggagatg gtgcgctcct ggacgtagcc 2581 ttcgggcatg gcggacttga agaagtcgtg ccgcttcatg tggtcggggt agcgggcgaa 2641 gcactgcacg ccccagctca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc 2701 ggtggtgcag atgaacttca gggtcagctt gccgatggtg gcatcgccct cgccctcgcc 2761 cctcacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac 2821 cccggtgaac agctcctcgc ccttgctgcc tccgaccggt gcgtcgccgg tgctgccggg 2881 cacccagagc agcagcaccc atagcaggag ggtgtcggtc tccatggtgg ctagaataac 2941 ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt 3001 ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcc 3061 tgcagccccg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 3121 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 3181 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 3241 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 3301 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 3361 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 3421 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt 3481 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 3541 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 3601 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 3661 catcgggggg atcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 3721 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 3781 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 3841 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggcg 3901 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac 3961 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag 4021 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct 4081 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca 4141 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 4201 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 4261 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 4321 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt 4381 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 4441 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg 4501 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 4561 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 4621 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 4681 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 4741 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt 4801 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 4861 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 4921 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 4981 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 5041 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 5101 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 5161 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 5221 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 5281 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 5341 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 5401 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 5461 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 5521 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 5581 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 5641 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 5701 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 5761 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 5821 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 5881 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 5941 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 6001 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 6061 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 6121 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac 6181 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 6241 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 6301 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 6361 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 6421 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 6481 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 6541 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 6601 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt //