LOCUS Exported 5914 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION FRET probe; extracelllular synapse targeted mScarlet. ACCESSION . VERSION . KEYWORDS pZac2.1 hSynapsin1 NAPA-N2 SV40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5914) AUTHORS Octeau JC, Chai H, Jiang R, Bonanno SL, Martin KC, Khakh BS TITLE An Optical Neuron-Astrocyte Proximity Assay at Synaptic Distance Scales. JOURNAL Neuron. 2018 Apr 4;98(1):49-66.e9. doi: 10.1016/j.neuron.2018.03.003. PUBMED 29621490 REFERENCE 2 (bases 1 to 5914) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5914 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 117..138 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 153..183 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 191..207 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 196..218 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 215..231 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 215..231 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" repeat_region 269..398 /label=AAV2 ITR promoter 475..922 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" intron 969..1101 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 1084..1103 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 1146..1164 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1269..1964 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(1402..1420) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1670..1689 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1965..2030 /label=MCS /note="multiple cloning site" polyA_signal 2949..3070 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2960..2979) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3014..3033 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 3232..3361 /label=AAV2 ITR primer_bind complement(3380..3397) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3380..3396) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3389..3411) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3538..3993 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3625..3644) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3835..3856 /label=F1ori-F /note="F1 origin, forward primer" promoter 4019..4123 /gene="bla" /label=AmpR promoter CDS 4124..4984 /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(4342..4361) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5155..5743 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5644..5663 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5897..5914 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 61 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat 121 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg 181 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac 241 gccagattta attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg 301 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg 361 cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc 421 atgctactta tctacgtagc catgctctag gaagatctga gggccctgcg tatgagtgca 481 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 541 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 601 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 661 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 721 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 781 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 841 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 901 cgtgtcgtgc ctgagagcgc agtcgagaga acttaagctg cagaagttgg tcgtgaggca 961 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct 1021 tgtcgagaca gagaaaactc ttgcgtttct gataggcacc tattggtctt actgacatcc 1081 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga 1141 gtacttaata cgactcacta taggctagcc tcgagaccac catgccacct tctactagtc 1201 ttttgttgct tgctgcactt ctccccttcg ctcttcccgc ttcagattgg aagacaggag 1261 aagtcaccat ggtgagcaag ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc 1321 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 1381 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct 1441 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcacc aagcaccccg 1501 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga 1561 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca 1621 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc 1681 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc 1741 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact 1801 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc 1861 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct 1921 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc 1981 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta 2041 gagtgtacaa gggcggtggc ggatctggcg gcggtggtag cgtcgacttt cagaaagcta 2101 gctctattgt ttataagaag gagggggaac aggtggagtt cagtttccca ttggccttca 2161 cagtcgagaa gctcacagga tcaggagagt tgtggtggca ggccgaaaga gcctcctcca 2221 gcaagagttg gatcactttt gatctgaaaa acaaagaagt gagcgtgaag agagtcacac 2281 aggaccctaa gctgcagatg ggcaagaagt tgcctttgca cttgacactc ccccaagccc 2341 tgccccagta tgctggctcc gggaacctca ccctcgcctt ggaggctaag accggaaagt 2401 tgcatcagga ggtcaacctc gtggtgatga gggcaacaca gctgcagaag aacctgacct 2461 gcgaagtgtg gggtcccacc tcacctaaac tgatgctgtc tctgaagctg gagaataagg 2521 aggctaaagt cagtaaaaga gagaaggctg tgtgggtcct taacccggag gcagggatgt 2581 ggcagtgtct tcttagcgac tctggacagg tgctgctgga gagcaacatc aaggtgcttc 2641 ctacttggtc aacaccagta cagccgatgg ccctgatcgt actcggggga gtcgccggcc 2701 tgcttctgtt tatcggcctg ggcatcttct tctgcgtacg ctacaagtat cgcaacaggg 2761 atgagggatc ttaccacgtt gacgaatcta ggaactatat cagtaacagc gctcagagta 2821 atggcgccgt ggttaaagag aaacagccct ccagcgccaa atcagcaaac aaaaacaaga 2881 aaaacaagga taaagagtac tacgtataat ctagagtcga cccgggcggc cgcttcgagc 2941 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa 3001 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa 3061 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg 3121 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc 3181 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc 3241 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac 3301 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca 3361 gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg 3421 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg 3481 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg 3541 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 3601 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 3661 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 3721 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 3841 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 3901 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 3961 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg 4021 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 4081 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt 4141 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 4201 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga 4261 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat 4321 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca 4381 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt 4441 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 4501 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct 4561 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 4621 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac 4681 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat 4741 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 4801 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 4861 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 4921 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 4981 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 5041 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 5101 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 5161 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 5221 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 5281 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa 5341 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 5401 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 5461 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 5521 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 5581 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 5641 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 5701 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 5761 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 5821 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag 5881 ccgaacgacc gagcgcagcg agtcagtgag cgag //