LOCUS Exported 6480 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Global labeling of inhibitory synapses (red fluorescence). ACCESSION . VERSION . KEYWORDS AAV-mRuby2-Gephyrin.FingR-IL2RGTC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6480) 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 6480) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6480 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 51..1229 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 282..1220 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1177..1197 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 1246..1265 /label=T7 /note="T7 promoter, forward primer" promoter 1246..1264 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 1291..1300 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1297..2007 /codon_start=1 /product="monomeric red fluorescent protein derived from mRuby, with improved photophysical properties (Lam et al., 2012)" /label=mRuby2 /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV AKFAGLGGGMDELYK" CDS 2314..2337 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 2755..2880 /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 2899..3487 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2952..2972) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3370..3381) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3490..3506) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3491..3507) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 3536..3657 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3547..3566) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3601..3620 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3865..3882) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4036..4624) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4116..4135) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4795..5655) /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 5418..5437 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5656..5760) /gene="bla" /label=AmpR promoter rep_origin complement(5786..6241) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5923..5944) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6135..6154 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 6313..6442 /label=AAV2 ITR ORIGIN 1 tagccatgct ctaggaagat cacgcgtctt ccacagagtg aattccgtga ggctccggtg 61 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 121 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 181 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 241 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 301 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 361 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 421 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 481 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 541 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 601 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 661 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 721 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 781 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 841 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 901 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 961 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1021 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1081 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1141 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1201 gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata cgactcacta 1261 tagggagacc caagctggct aggtaagctt gccaccatgg tgtctaaggg cgaagagctg 1321 atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc 1381 aaatgcacag gtgaaggaga aggcaatccg tacatgggaa ctcaaaccat gaggatcaaa 1441 gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat 1501 ggcagccgta cttttatcaa gtacccgaaa ggcattcctg atttctttaa acagtccttt 1561 cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc 1621 atgcaggaca ccagccttga ggatggctgt ctcgtttacc acgtccaagt cagaggggta 1681 aactttccct ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca 1741 gagatgatgt atccagcaga tggtggtctg aggggataca ctcatatggc actgaaagtt 1801 gatggtggtg gccatctgtc ttgctctttc gtaacaactt acaggtcaaa aaagaccgtc 1861 gggaacatca agatgcccgg tatccatgcc gttgatcacc gcctggaaag gttagaggaa 1921 agtgacaatg aaatgttcgt agtacaacgc gaacacgcag ttgccaagtt cgccgggctt 1981 ggtggtggga tggacgagct gtacaagtct aggggagctg gcgctggagc gggtgcaggg 2041 gctggcctcg aagtcaagga agcatcacca accagcatcc agatcagctg gggcaagtac 2101 aaggtcatgg ttcgctacta ccgcatcacc tacggtgaaa ctggtggcaa tagccctgtc 2161 caggaattca ccgtgcctgg cagcaagtcc actgctacca tcagcagcct gaaacctggt 2221 gtcgactata ccatcacggt gtacgccgtc acgatcgacc actggaacta ccaggacccg 2281 atcccgatct ccatcaacta ccgcaccgga tccgattaca aggatgacga cgataagggt 2341 agcggctcca gtagatctgg gtttcagtgc cagatttgca tgcgcaactt tagccgcaaa 2401 agcaccctga ccgatcatat tcgcacccat accggcgaaa aaccgtttgc gtgcgatatt 2461 tgcggccgca aatttgcggc gcgcagcacc cgcaccaccc ataccaaaat tcataccggc 2521 agccagaaac cgtttcagtg ccgcatttgc atgcgcaact ttagccgcag cgatagcctg 2581 agcaaacata ttcgcaccca taccggcgaa aaaccgtttg cgtgcgatat ttgcggccgc 2641 aaatttgcgc agcgcagcaa cctgaaagtg cataccaaaa ttcatctgcg cggcagccag 2701 ctgatcgatg gtgtcgacgc tcctgaacaa cgtgaaggtg cttctcaagt ttctgtttct 2761 gttacttttg aagatgttgc tgttcttttt actcgtgatg aatggaaaaa acttgatctt 2821 tctcaacgtt ctctttatcg tgaagttatg cttgaaaatt attctaatct tgcttctatg 2881 gcttaaaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt 2941 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 3001 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 3061 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 3121 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 3181 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 3241 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 3301 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 3361 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 3421 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 3481 tccccgcatc gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga 3541 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt 3601 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac 3661 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa 3721 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta 3781 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt 3841 ggccactccc tctctgyscg ctysctcgct cactgactcg ctgcgctcgg tcgttcggct 3901 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 3961 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 4021 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 4081 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 4141 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 4201 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 4261 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 4321 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 4381 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 4441 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 4501 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 4561 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 4621 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 4681 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 4741 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 4801 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 4861 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 4921 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 4981 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 5041 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 5101 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 5161 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 5221 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 5281 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 5341 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 5401 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 5461 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 5521 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 5581 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 5641 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 5701 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 5761 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg 5821 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct 5881 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt 5941 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat 6001 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca 6061 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac 6121 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta 6181 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg 6241 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 6301 ctattacgcc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt 6361 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc 6421 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg //