LOCUS Exported 6465 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Global labeling of inhibitory synapses (red fluorescence). ACCESSION . VERSION . KEYWORDS AAV-EF1A-mScarlet-Gephyrin.FingR-IL2RGTC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6465) AUTHORS Bensussen S, Shankar S, Ching KH, Zemel D, Ta TL, Mount RA, Shroff SN, Gritton HJ, Fabris P, Vanbenschoten H, Beck C, Man HY, Han X TITLE A Viral Toolbox of Genetically Encoded Fluorescent Synaptic Tags. JOURNAL iScience. 2020 Jun 30;23(7):101330. doi: 10.1016/j.isci.2020.101330. PUBMED 32674057 REFERENCE 2 (bases 1 to 6465) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6465 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR promoter 219..1397 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 450..1388 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1345..1365 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 1414..1433 /label=T7 /note="T7 promoter, forward primer" promoter 1414..1432 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 1459..1468 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1465..2160 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet-I /note="matures faster than mScarlet" /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(1598..1616) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1866..1885 /label=mCherry-F /note="mCherry, forward primer" CDS 2467..2490 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 2908..3033 /codon_start=1 /product="Kruppel-associated box (KRAB) transcriptional repression domain from the rat zinc finger protein Kid-1 (Witzgall et al., 1994)" /label=KRAB-A /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM" misc_feature 3052..3640 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3105..3125) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3523..3534) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3643..3659) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3644..3660) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 3689..3810 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3700..3719) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3754..3773 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(4018..4035) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4189..4777) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4269..4288) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4948..5808) /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 5571..5590 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5809..5913) /gene="bla" /label=AmpR promoter rep_origin complement(5939..6394) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6076..6097) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6288..6307 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct 181 aggaagatca cgcgtcttcc acagagtgaa ttccgtgagg ctccggtgcc cgtcagtggg 241 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg 301 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc 361 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt 421 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg 481 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg 541 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt 601 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg 661 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt 721 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg 781 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg 841 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac 901 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg 961 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc 1021 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg 1081 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg 1141 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta 1201 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg 1261 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt 1321 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc 1381 catttcaggt gtcgtgagga attagcttgg tactaatacg actcactata gggagaccca 1441 agctggctag gtaagcttgc caccatggtg agcaagggcg aggcagtgat caaggagttc 1501 atgcggttca aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc 1561 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt 1621 ggccccctgc ccttctcctg ggacatcctg tcccctcagt tcatgtacgg ctccagggcc 1681 ttcatcaagc accccgccga catccccgac tactataagc agtccttccc cgagggcttc 1741 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc 1801 tccctggagg acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct 1861 gacggccccg taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac 1921 cccgaggacg gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc 1981 cgctacctgg cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc 2041 gcctacaacg tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg 2101 gaacagtacg aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag 2161 tctaggggag ctggcgctgg agcgggtgca ggggctggcc tcgaagtcaa ggaagcatca 2221 ccaaccagca tccagatcag ctggggcaag tacaaggtca tggttcgcta ctaccgcatc 2281 acctacggtg aaactggtgg caatagccct gtccaggaat tcaccgtgcc tggcagcaag 2341 tccactgcta ccatcagcag cctgaaacct ggtgtcgact ataccatcac ggtgtacgcc 2401 gtcacgatcg accactggaa ctaccaggac ccgatcccga tctccatcaa ctaccgcacc 2461 ggatccgatt acaaggatga cgacgataag ggtagcggct ccagtagatc tgggtttcag 2521 tgccagattt gcatgcgcaa ctttagccgc aaaagcaccc tgaccgatca tattcgcacc 2581 cataccggcg aaaaaccgtt tgcgtgcgat atttgcggcc gcaaatttgc ggcgcgcagc 2641 acccgcacca cccataccaa aattcatacc ggcagccaga aaccgtttca gtgccgcatt 2701 tgcatgcgca actttagccg cagcgatagc ctgagcaaac atattcgcac ccataccggc 2761 gaaaaaccgt ttgcgtgcga tatttgcggc cgcaaatttg cgcagcgcag caacctgaaa 2821 gtgcatacca aaattcatct gcgcggcagc cagctgatcg atggtgtcga cgctcctgaa 2881 caacgtgaag gtgcttctca agtttctgtt tctgttactt ttgaagatgt tgctgttctt 2941 tttactcgtg atgaatggaa aaaacttgat ctttctcaac gttctcttta tcgtgaagtt 3001 atgcttgaaa attattctaa tcttgcttct atggcttaaa agcttatcga taatcaacct 3061 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 3121 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 3181 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 3241 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 3301 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 3361 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 3421 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 3481 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 3541 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 3601 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga 3661 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca 3721 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt 3781 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt 3841 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt 3901 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat 3961 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctysctc 4021 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 4081 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 4141 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 4201 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 4261 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 4321 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 4381 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 4441 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 4501 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 4561 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 4621 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 4681 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 4741 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 4801 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 4861 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 4921 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 4981 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 5041 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 5101 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 5161 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 5221 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 5281 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 5341 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 5401 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 5461 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 5521 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 5581 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 5641 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 5701 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 5761 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 5821 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 5881 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa 5941 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt 6001 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata 6061 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac 6121 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa 6181 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc 6241 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg 6301 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca 6361 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac 6421 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccag //