LOCUS Exported 5117 bp ds-DNA circular SYN 21-JUL-2022 DEFINITION Donor plasmid to endogenously tag GLE gene in Drosophila melanogaster ovarian somatic sheet (OSS) cells. . ACCESSION . VERSION . KEYWORDS pDonor_N_FH_GLE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5117) AUTHORS Meng Q, Stoyko D, Andrews CM, Konstantinidou P, Genzor P, O T, Elchert AR, Benner L, Sobti S, Katz EY, Haase AD TITLE Functional editing of endogenous genes through rapid selection of cell pools (Rapid generation of endogenously tagged genes in Drosophila ovarian somatic sheath cells). JOURNAL Nucleic Acids Res. 2022 May 27. pii: 6594084. doi: 10.1093/nar/gkac448. PUBMED 35639929 REFERENCE 2 (bases 1 to 5117) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 21, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5117 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 118..137 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 371..388 /label=L4440 /note="L4440 vector, forward primer" protein_bind 505..526 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 541..571 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 579..595 /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 584..606 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 603..619 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 603..619 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 1241..1375 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1265..1284) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1319..1338 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(1376..1975) /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1459..1479) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 1956..1975 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" CDS 2330..2395 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 2432..2521 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA" primer_bind complement(3120..3137) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3120..3136) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3129..3151) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(3345..3364) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3464..3486 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3524..3542) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3610..3714 /gene="bla" /label=AmpR promoter CDS 3715..4575 /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(3933..3952) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4746..217 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 601 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg ggtctacgga 661 gctgagtatc tttgtgagaa aactaccgaa gaagaaggca gttatggctg tgaagaaagt 721 cactaaatta tttaatcgcc tcatttatgt gttgttttga ctagagagac cgtgaaacat 781 tcgcgaaaat ataccatgtt tactggggta tcgcaaattt gttcattgaa tctcttgctt 841 gcattgtgaa taatcgttaa taattgaata atcgttaatt ttggttttat cataaagtgg 901 aaagcaaatt atatagactg tatacttatg tatactgtct taaaagtaag ttaaataaag 961 tttgcactaa taatatactt tacaaattcg agttcggaat cgctggacca aagcatttca 1021 attcgtaagt aagaccgcaa gtaagaccgt atccccttaa caataacgga atcctgattg 1081 ccgcccaatg cgagtaatat taaattgagt tggcagtctt atgctatatt tttgttgatt 1141 aaaaagtgta aaaacatatt cgaaattaaa agtagtaatc gtattaagac actgggttgc 1201 gattggcgct attcgagacc ttttagagcc ccaggtaagt gatccagaca tgataagata 1261 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga 1321 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttctagg 1381 caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg 1441 tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga 1501 aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca 1561 gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct 1621 tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc 1681 tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg 1741 gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga 1801 gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg 1861 ggtcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg 1921 cccgggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcattttga 1981 ttgtctctgg attagacgac tgctggctga tggagcggct ttgtgtcggg aggagtatcc 2041 acacagcaca agaactcaaa cggtagtgat atgaaaactg gtcccgcagc ccgttttata 2101 aagcccgctt agcgctaaaa ccttatcagc cgcaatgctc tccgctctca agtcgcgttc 2161 aaaactttta ccatatatgg gtatgaattg ttcgccgatt gggttttcat ttacggagaa 2221 tttcctccgc aactgggtgt tcgtgcttgt gtgtgtcttg tatatcgcca tgaagagtgt 2281 acctaatttt aatcaaccga tttctctctc ttccgcagga atcaaaatgg actacaaaga 2341 ccatgacggt gattataaag atcatgacat cgattacaag gatgacgatg acaagggtgg 2401 atctggaggt tccggcggct cagggggtag ttacccatac gatgttcctg actatgcggg 2461 ctatccctat gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc 2521 ttccggtggc tctggctctt caggctcaat ggacgacatg atgcgggcga tggactgtct 2581 aaatatggca gcgacgttgc gcaacgcggc tctggcgtcc gcaacgtgca cgggcaggac 2641 tttgggcgag gatcgggaac ctatatgggt ggagggcagt cggaagacac ctgagccacc 2701 attacccgaa gaaagcccag ctccagagcc caataacgag attcccctgc tgcccaaaat 2761 agattttgag cctaatatat cctgctttcc cgacctacag gctatcaacg ctagcattgt 2821 taggcgggaa ctggaggagg agggtaagag gtgtgtccgc cagcagctaa aggcgataag 2881 ggacaaacag gacgccatgc gcctgtcgcg agagacgcag caaaggaagg aggaacgcca 2941 gcgcgaccag ttgcaacaaa aggcgctgcg ggaacgcaat gaaagtctcc ttatccagaa 3001 agctgaccag atgactgcgg cacaactaga ggcccagcag cgcgagcaat tggcactgcg 3061 ccaacagatt gaccagaagc tgcacaagtt gaggatcccc gggtaccgag ctcgaattca 3121 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 3181 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 3241 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt 3301 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat 3361 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct 3421 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt 3481 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta 3541 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg 3601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 3661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 3721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 3781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 3841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 3901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 3961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 4021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 4081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 4141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 4201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 4261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 4321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 4381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 4441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 4501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 4561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 4621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 4681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 4741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 4801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 4861 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 4921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 4981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 5041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 5101 acgacctaca ccgaact //