LOCUS Exported 7444 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For membrane contact. ACCESSION . VERSION . KEYWORDS Prig-3::CD4::spGFP11 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7444) AUTHORS Feinberg EH, Vanhoven MK, Bendesky A, Wang G, Fetter RD, Shen K, Bargmann CI TITLE GFP Reconstitution Across Synaptic Partners (GRASP) defines cell contacts and synapses in living nervous systems. JOURNAL Neuron. 2008 Feb 7. 57(3):353-63. PUBMED 18255029 REFERENCE 2 (bases 1 to 7444) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7444 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 3139..3186 /codon_start=1 /product="11th beta-strand of superfolder GFP" /label=GFP11 /note="associates with GFP(1-10), consisting of the first 10 beta-strands, to reconstitute fluorescent GFP (Cabantous et al., 2005; Kamiyama et al., 2016)" /translation="RDHMVLHEYVNAAGIT" primer_bind complement(5013..5035) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5135..5154 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5236..5254) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5322..5426 /gene="bla" /label=AmpR promoter CDS 5427..6287 /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(5645..5664) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6458..7046 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6947..6966 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7200..7217 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7334..7355 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7370..7400 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7408..7424 /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 7413..7435 /label=M13/pUC Reverse /note="In lacZ gene" ORIGIN 1 atgaccatga ttacgccaag cttgcatgcg cggccgcact gactgggccg gccgaaatgc 61 gacccatcat attgttccta ttgtcatgta caagctgtcc ctgttttgct cgcttaatcc 121 gtcaacccct tccccgttct aattagagat gagtgaatta caatagagtt gaaacggaaa 181 tctgagtgtc tgtagctttc cgccgccgct tttgttacga gatggtagac gtccctttct 241 tggtgtttat tgtctcacaa actgtataat aaagcaccgt tgtgtaaaag gcatttggta 301 tcaggaagat aaaatgaata aaatgcaaat gcggaatcaa aaattgccaa ataattttga 361 ccaataaaat atcaaaaata atagaacaga aataaacaaa taaaactacg gagaaaatgt 421 gattattcca ttaaacaacg ttttaaacca actttattcg tcagtgggga aggttttgga 481 caaatatcat agtaaaactt gttcgtttta attatacata atcaataatt ataggtgtta 541 gaaccaatgg tcgaaattta aaaaaaaatc aacattttag aaaatttcgg caatttaccg 601 aaatattgac tgacaatttt acatatataa gtgttttgaa atagaatccc cattgacgat 661 gcgccacctt aaacagtaga actgttccaa tcagaaacta aataaatacg gttattatat 721 atatatatat atatatatat atataacttg tttttattat gcatttctca aaactcacca 781 aactcaatct ctaatttacg gttgttgagg aaaatatatt tatcacattt agttgagact 841 ctaaaagaag tacaactttt ttcttgcaat tcctccacca atccaatttt tcaagagtca 901 attgcttgga gtcctcgttt tctcgtaatc cgatattccc cctatttctg tgattttatt 961 cagtttcgtc ctcttacacc aaaaatcttc atgacttttt tggttttctg cttaatcttt 1021 tttttgtttt taaagtctac catccggcaa ttttttttgc aaatggttgc tcagaatttg 1081 acctcagatt aaaattttca ggaacttcct ggtcccattt cgtcacattt aataacaaac 1141 caaatccttc agcttgagtc gtccaagcca acgcaaccct aattcgattc cacctccaat 1201 ttaccttttt gccgttttcg atcaaattgt gggggctctt tttatttgaa agtcgtaaaa 1261 tggcgcgtcc ttgtcgaaaa ttgagatagc taatggttcc aaataaaatc aaatgcatga 1321 acctagtagg aaaaatcgtt ttttttttca tcttccaaag aaaaagttta agataaaccc 1381 accactccag caatccacct acgcgttttt caccggaaac tgtcatttgt ttttgatgta 1441 tgacagtgcc agcactgaac taaattccca tttgttttgt tgtggtgtta tctaaccacg 1501 tggctctttc tctaaatttt gcgtaggctg caagatccgc aatcccacca aaaaatggtt 1561 tctaatgtac ttcatacata tcccacatga ctcagtgctc cgaaacgccc agcaaccccc 1621 gcgtttgtct ttacaaaaat gaggcggaaa tgggaaattt tcctgctatt gctatggtaa 1681 aatacttaag ctgtattgag atttgggaaa ttgaaagata agatgaaaaa tccactctgt 1741 ataattaaaa aggtttattt tttttgtaat ttttgtaccg caaatttttt cattaactga 1801 tttttcttta ttcattcgat ggcccagttt cagatcttca ttatcatgta gtttattcat 1861 tcaaactatt tgtgacatca tgatggttgt ttaaaaataa tttcgaatga agaaagctta 1921 aacctattat aattttcttt tctataaaaa atattgattt tgtaaagcgt ttggcagtga 1981 cagtaccaaa aaatgtttta aaatgattta agaaaatttt ttattttatt tgtataatac 2041 gcgtaaaatg agtatactcg tcatttgtca cgaatcgagt tctcagtttg ctgtgaatat 2101 cttgtaatag aaatcgatct catagtgtca cttttgaggc ataacttttc aggttgttac 2161 aagttcatta acttgtcaag actgggtcat gttttaaagt gtaactgtta cccatgtacc 2221 ctaaactttg agacaaactc ctcaaaactt gtaaccagaa tcctatgtag atgactcaca 2281 aaaccctgtt ttcaatttga acgaatttcg aaactggcca aaatgttccc ctcttgctca 2341 cctaatgctt ttaccattta aaaaggatgg taacaacact ctttcgaaaa tgtaggcggt 2401 ccattttcgg tttctcataa ccgtctggct ctatctccta gtgcgcgagg agagggcggg 2461 cggagtagga atcgggggca gcgcactcat acactctgcc cttctctttt gcacacacag 2521 aaaaagatag atacagacca acacttattg ttgtccagta agcttccaga aaactatttt 2581 cagaattaca aactacattt ttctagaaca gtcaattgat tttttacaag attcgagtat 2641 tctaacatta gtgaaagtat ttagccactg caattttcaa ctgtaggttt caacttccaa 2701 aaaaaattcc tagcatttct aattggatga aaaaaggcta tctagaccct ccattattta 2761 ctcataattt catctcgtcc tcatttttaa tcccctcatt gatcccatca caatccgcca 2821 aaatgttttt taaacacttt tttcttcacc agccgtatgt ggtcaactgc ctaaaaattt 2881 cacagctatt tgtcattcct tgcagaagaa cttcattctt ctttttcgaa aggcgcgcct 2941 ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg atactaacat 3001 aacatagaac attttcagga ggacccttgg agggtaccgc cggcaatgcc accttcaaca 3061 tcattgctgc tcctcgcagc acttcttcca ttcgctttac cagcaagcga ttggaagact 3121 ggagaagtca ctgctagcag agatcatatg gtgttgcatg aatatgtcaa tgctgcaggt 3181 attactggag gaggaggatc aggaggtgga ggatctttcc aaaaagcatc atctattgtt 3241 tataagaagg aaggtgagta agtttaaaca gatccatact aactaacttg ttctgacata 3301 attttcagac aagtcgagtt ttcattcccg cttgccttta ctgttgagaa acttactgga 3361 tctggagaac tttggtggca agccgaaaga gcttcttcat caaagtcatg gattactttt 3421 gatttgaaaa ataaagaggt aagtttaaac agttgaatac taactaacgg agatctttga 3481 aattttcagg tgtcagtgaa aagagtgaca caagatccga aattgcagat gggaaagaaa 3541 cttccacttc atttgacttt gccacaagcc ttgccacagt atgcaggttc aggaaatttg 3601 acacttgctt tggaagccaa gtaagtttaa acaataacct actaactaac gtagataatt 3661 taaattttca gaacaggaaa gcttcaccaa gaggtcaatc ttgtggtgat gcgagctact 3721 caattgcaga agaatcttac atgcgaagtt tggggaccaa catctccaaa attgatgttg 3781 tcattgaaac ttgaaaacaa gtaagtttaa acttgatagt actaactaac atgtttcatt 3841 taaattttca gggaggcaaa agtgtctaag cgtgagaaag ctgtttgggt tcttaaccca 3901 gaagcaggaa tgtggcagtg tttgttgtca gattctggtc aagtgttgct tgaatcaaat 3961 attaaggtct tgccgacatg ggtaagttta aactgagttc tactaactaa cgagtaatat 4021 ttaaattttc agtctacacc ggttcagcca atggcactta ttgttcttgg tggtgtggca 4081 ggacttttgt tgttcatcgg attgggaatc ttcttctgtg tccgatgccg acatcgacgt 4141 cgttaaggta ccgacgtcat gcgaagggcg aattctgcag atatctgagc tccgcatcgg 4201 ccgctgtcat cagatcgcca tctcgcgccc gtgcctctga cttctaagtc caattactct 4261 tcaacatccc tacatgctct ttctccctgt gctcccaccc cctatttttg ttattatcaa 4321 aaaaacttct tcttaatttc tttgtttttt agcttctttt aagtcacctc taacaatgaa 4381 attgtgtaga ttcaaaaata gaattaattc gtaataaaaa gtcgaaaaaa attgtgctcc 4441 ctccccccat taataataat tctatcccaa aatctacaca atgttctgtg tacacttctt 4501 atgttttttt tacttctgat aaattttttt tgaaacatca tagaaaaaac cgcacacaaa 4561 ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct tcgcacgtct 4621 gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta tttttggaat 4681 ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt tgggggtttc 4741 ccctattgtt tgtcaagagt ttcgaggacg gcgtttttct tgctaaaatc acaagtattg 4801 atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga 4861 gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag 4921 aatacattcc caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt 4981 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg 5041 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg 5101 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga 5161 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg 5221 cggccttaat taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa 5281 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 5341 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 5401 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 5461 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 5521 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 5581 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 5641 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 5701 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 5761 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 5821 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 5881 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 5941 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 6001 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 6061 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 6121 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 6181 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 6241 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 6301 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 6361 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 6421 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 6481 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 6541 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 6601 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 6661 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 6721 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 6781 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 6841 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 6901 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 6961 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 7021 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 7081 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 7141 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 7201 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc 7261 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 7321 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta 7381 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 7441 gctn 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