LOCUS SuRe.AD1(TP901-1 5724 bp DNA circular SYN 26-APR-2025 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5724) AUTHORS Junjie Luo TITLE Direct Submission JOURNAL Exported Apr 26, 2025 from SnapGene 8.0.3 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5724 /mol_type="other DNA" /note="color: #ffffff" /organism="synthetic DNA construct" source 99..108 /chromosome="2R" /mol_type="genomic DNA" /note="/genotype=y[1]; Gr22b[1] Gr22d[1] cn[1] CG33964[R4.2]bw[1] sp[1]; LysC[1] MstProx[1] GstD5[1] Rh6[1]" /note="color: #ffffff" /db_xref="taxon:7227" /organism="Drosophila melanogaster" source 129..138 /chromosome="2R" /mol_type="genomic DNA" /note="/genotype=y[1]; Gr22b[1] Gr22d[1] cn[1] CG33964[R4.2]bw[1] sp[1]; LysC[1] MstProx[1] GstD5[1] Rh6[1]" /note="color: #ffffff" /db_xref="taxon:7227" /organism="Drosophila melanogaster" source 159..168 /chromosome="2R" /mol_type="genomic DNA" /note="/genotype=y[1]; Gr22b[1] Gr22d[1] cn[1] CG33964[R4.2]bw[1] sp[1]; LysC[1] MstProx[1] GstD5[1] Rh6[1]" /note="color: #ffffff" /db_xref="taxon:7227" /organism="Drosophila melanogaster" source 3263..3272 /chromosome="2R" /mol_type="genomic DNA" /note="/genotype=y[1]; Gr22b[1] Gr22d[1] cn[1] CG33964[R4.2]bw[1] sp[1]; LysC[1] MstProx[1] GstD5[1] Rh6[1]" /note="color: #ffffff" /db_xref="taxon:7227" /organism="Drosophila melanogaster" source 3293..3302 /chromosome="2R" /mol_type="genomic DNA" /note="/genotype=y[1]; Gr22b[1] Gr22d[1] cn[1] CG33964[R4.2]bw[1] sp[1]; LysC[1] MstProx[1] GstD5[1] Rh6[1]" /note="color: #ffffff" /db_xref="taxon:7227" /organism="Drosophila melanogaster" source 3323..3332 /chromosome="2R" /mol_type="genomic DNA" /note="/genotype=y[1]; Gr22b[1] Gr22d[1] cn[1] CG33964[R4.2]bw[1] sp[1]; LysC[1] MstProx[1] GstD5[1] Rh6[1]" /note="color: #ffffff" /db_xref="taxon:7227" /organism="Drosophila melanogaster" misc_feature 64..181 /label=3xTpnC41C_PCR /note="color: #a6acb3" misc_feature join(81..96,99..108,111..126,129..138,141..156,159..168) /label=3xTpnC41C /note="Expression of the Troponin C at 41C Gene in Adult Drosophila Tubular Muscles Depends upon Both Positive and Negative Regulatory Inputs Maria B. Chechenova, Sara Maes, Richard M. Cripps Published: December 7, 2015 '>https://doi.org/10.1371/journal.pone.0144615" /note="This forward directional feature has 6 segments: 1: 81 .. 96 / #a6acb3 / SM3 2: 99 .. 108 / #ff00ff / MEF2 3: 111 .. 126 / #a6acb3 / SM3 4: 129 .. 138 / #ff00ff / MEF2 5: 141 .. 156 / #a6acb3 / SM3 6: 159 .. 168 / #ff00ff / MEF2" promoter join(209..214,215..220,248..333) /label=hsp70 mini /note="Drosophila melanogaster hsp70Bb promoter" /note="This forward directional feature has 3 segments: 1: 209 .. 214 / #ffffff 2: 215 .. 220 / #cc99ff / TATA box 3: 248 .. 333 / #ffffff / align to 3xP3" regulatory 347..356 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /note="color: #a6acb3" /regulatory_class="other" CDS join(353..355,356..358,359..1072) /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /note="This feature has 3 segments: 1: 353 .. 355 / #05fd14 2: 356 .. 358 / #05fd14 / 1a 3: 359 .. 1072 / #05fd14" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" polyA_signal 1213..1334 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" promoter 1377..1797 /label=dU6-3 promoter /note="RNA polymerase III promoter for Drosophila U6-3 snRNA (Port et al., 2014)" /note="color: #ffffff; direction: RIGHT" misc_feature 1800..1819 /label=gRNAu /label=attB_gRNA(core) /note="gRNAu (targets Janelia-based vector seq*) - ACCGTCGACGATGTAGGTCANGG " /note="color: #00ff00; direction: RIGHT" misc_RNA 1820..1905 /label=LgRNA scaffold /note="long gRNA, removed tttt, extended loop guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" /note="color: #00ccff; direction: RIGHT" primer_bind complement(1934..1950) /label=SK primer /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" misc_feature complement(1967..2254) /label=PhiC31 attB /note="color: #a6acb3; direction: LEFT" misc_feature complement(1975..2254) /label=attB /note="color: #a6acb3; direction: LEFT" protein_bind complement(2169..2238) /label=attB /bound_moiety="phage phi-C31 integrase" /note="attB site for the phi-C31 integrase (Groth et al., 2000)" /note="color: #31849b; direction: LEFT" misc_feature complement(join(2240..2242,2243..2245,3912..3928)) /label=gRNAu target with AD1(TP901-1)1 insertion /label=attB_SgrDI_gRNA /note="This reverse directional feature has 3 segments: 1: 2240 .. 2242 / #00ff00 2: 2243 .. 2245 / #00ff00 3: 3912 .. 3928 / #00ff00" misc_feature complement(join(2240..2242,2246..2254)) /label=attB_SgrDI_gRNA /note="This reverse directional feature has 2 segments: 1: 2240 .. 2242 / #00ff00 2: 2246 .. 2254 / #00ff00" misc_feature join(2247..2263,2264..3350,3351..3372,3373..3505,3506..3512, 3513..3898,3899..3911) /label=[AD1_P^{TP901}] /note="This forward directional feature has 7 segments: 1: 2247 .. 2263 / #ffffff 2: 2264 .. 3350 / #ff0000 / TpnC41C-mCherry 3: 3351 .. 3372 / #ffffff 4: 3373 .. 3505 / #ff00ff / attP[TP901] 5: 3506 .. 3512 / #ffffff 6: 3513 .. 3898 / #31849b / Insulator 7: 3899 .. 3911 / #ffffff" polyA_signal 2269..2390 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" CDS complement(2397..3107) /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /note="color: #ff0000" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" promoter complement(join(3118..3203,3231..3236,3237..3242)) /label=hsp70 mini /note="Drosophila melanogaster hsp70Bb promoter" /note="This reverse directional feature has 3 segments: 1: 3118 .. 3203 / #ffffff / align to 3xP3 2: 3231 .. 3236 / #cc99ff / TATA box 3: 3237 .. 3242 / #ffffff" misc_feature 3250..3367 /label=3xTpnC41C_PCR /note="color: #a6acb3" misc_feature complement(join(3263..3272,3275..3290,3293..3302,3305..3320, 3323..3332,3335..3350)) /label=3xTpnC41C /note="Expression of the Troponin C at 41C Gene in Adult Drosophila Tubular Muscles Depends upon Both Positive and Negative Regulatory Inputs Maria B. Chechenova, Sara Maes, Richard M. Cripps Published: December 7, 2015 '>https://doi.org/10.1371/journal.pone.0144615" /note="This reverse directional feature has 6 segments: 1: 3263 .. 3272 / #ff00ff / MEF2 2: 3275 .. 3290 / #a6acb3 / SM3 3: 3293 .. 3302 / #ff00ff / MEF2 4: 3305 .. 3320 / #a6acb3 / SM3 5: 3323 .. 3332 / #ff00ff / MEF2 6: 3335 .. 3350 / #a6acb3 / SM3" misc_feature complement(3373..3505) /label=attP(TP901-1)Long /note="Blast by attP(TP901-1). choose the common long region" /note="color: #ff00ff; direction: LEFT" protein_bind 3513..3898 /label=gypsy insulator /bound_moiety="Su(Hw), CP190, andMod(mdg4)67.2" /note="chromatin insulator from Drosophila" /note="color: #31849b" rep_origin complement(3991..4579) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(join(4750..5541,5542..5610)) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 4750 .. 5541 / #ccffcc 2: 5542 .. 5610 / #ccffcc / signal sequence Cleavage site after base 5541" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5611..5715) /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: LEFT" ORIGIN 1 ttaattaact cgcgttaaga tacattgatg agtttggaca aacctgcagg ccctttcgtc 61 ttcctaactt ccgtgacccg ttcacaaata ccatttccct aaaaataacc ttcacaaata 121 ccatttccct aaaaataacc ttcacaaata ccatttccct aaaaataaag cggagactct 181 aagcttatcg atttcgaacc ctcgaccgcc ggagtataaa tagaggcgct tcgtctacgg 241 agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag ctaagcaaat 301 aaacaagcgc agctgaacaa gctaaacaat cggctcgaag ccggtcgcca ccatggtgag 361 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt 421 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct 481 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 541 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 601 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga 661 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 721 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 781 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa 841 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta 901 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag 961 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga 1021 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggcct 1081 aagcgatcgc tcaggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt 1141 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 1201 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 1261 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 1321 atcaatgtat cttacctgca ggagtaaact cgtcgacggt atcgataagc gctagctttt 1381 ttgctcacct gtgattgctc ctactcaaat acaaaaacat caaattttct gtcaataaag 1441 catatttatt tatatttatt ttacaggaaa gaattccttt taaagtgtat tttaacctat 1501 aatgaaaaac gattaaaaaa aatacataaa ataattcgaa aatttttgaa tagcccaggt 1561 tgataaaaat tcatttcata cgttttataa cttatgcccc taagtatttt ttgaccatag 1621 tgtttcaatt ctacattaat tttacagagt agaatgaaac gccacctact cagccaagag 1681 gcgaaaaggt tagctcgcca agcagagagg gcgccagtgc tcactacttt ttataattct 1741 caacttcttt ttccagactc agttcgtata tatagaccta ttttcaattt aacgtcggta 1801 ccgtcgacga tgtaggtcag tcttagagct atgctggaaa cagcatagca agttaagata 1861 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tatcgaattc 1921 ctgcagcccg ggggatccac tagttctaga gcggcgcgcc catatgttcg gcttgtcgac 1981 atgcccgccg tgaccgtcga gaacccgctg acgctgcccc gcgtatccgc acccgccgac 2041 gccgtcgcac gtcccgtgct caccgtgacc accgcgccca gcggtttcga gggcgagggc 2101 ttcccggtgc gccgcgcgtt cgccgggatc aactaccgcc acctcgaccc gttcatcatg 2161 atggaccaga tgggtgaggt ggagtacgcg cccggggagc ccaagggcac gccctggcac 2221 ccgcaccgcg gcttcgagac cgtgacctac atcgtctcga gacctaggta agatacattg 2281 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 2341 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt accggtttac 2401 ttgtacagct cgtccatgcc gccggtggag tggcggccct cggcgcgttc gtactgttcc 2461 acgatggtgt agtcctcgtt gtgggaggtg atgtccaact tgatgttgac gttgtaggcg 2521 ccgggcagct gcacgggctt cttggccttg taggtggtct tgacctcagc gtcgtagtgg 2581 ccgccgtcct tcagcttcag cctctgcttg atctcgccct tcagggcgcc gtcctcgggg 2641 tacatccgct cggaggaggc ctcccagccc atggttttct tctgcattac ggggccgtcg 2701 gaggggaagt tggtgccgcg cagcttcacc ttgtagatga actcgccgtc ctgcagggag 2761 gagtcctggg tcacggtcac cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg 2821 aagccctcgg ggaaggacag cttcaagtag tcggggatgt cggcggggtg cttcacgtag 2881 gccttggagc cgtacatgaa ctgaggggac aggatgtccc aggcgaaggg cagggggcca 2941 cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg 3001 ccctcgatct cgaactcgtg gccgttcacg gagccctcca tgtgcacctt gaagcgcatg 3061 aactccttga tgatggccat gttatcctcc tcgcccttgc tcaccatgtt gacgcgtccg 3121 attgtttagc ttgttcagct gcgcttgttt atttgcttag ctttcgctta gcgacgtgtt 3181 cactttgctt gtttgaattg aattgtcgct ccgtagacga agcgcctcta tttatactcc 3241 ggcggccgct agagtctccg ctttattttt agggaaatgg tatttgtgaa ggttattttt 3301 agggaaatgg tatttgtgaa ggttattttt agggaaatgg tatttgtgaa cgggtcacgg 3361 aagttagcat gctaactgat gaactatggg accaaataaa atatattgaa gaaattaatt 3421 gattacagaa actccttaaa aggagttttt tagttacctt aattgaaata aacgaaataa 3481 aaactcgcaa ttaagcgagt tggaagagct cgaattgatc ggctaaatgg tatggcaaga 3541 aaaggtatgc aatataataa tcttttattg ggtatgcaac gaaaatttgt ttcgtcaacg 3601 tatgcaatat tctttattaa aagagggtat gcaatgtatt ttattaaaaa cgggtatgca 3661 atataataat cttttattgg gtatgcaacg aaaatttgtt tcgtcaaagt atgcaatatt 3721 ttttattaaa agagggtatg caatgtattt tattaaaaac gggtatgcaa taaaaaatta 3781 tttggtttct ctaaaaagta tgcagcactt attttttgat aaggtatgca acaaaatttt 3841 actttgccga aaatatgcaa tgtttttgcg aataaattca acgcacactt attacgtggg 3901 cccacacgtg gcctacatcg tcgacggtaa catgtgagca aaaggccagc aaaaggccag 3961 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 4021 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 4081 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 4141 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 4201 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 4261 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 4321 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 4381 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 4441 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 4501 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 4561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 4621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 4681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 4741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 4801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 4861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 4921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 4981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 5041 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 5101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 5161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 5221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 5281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 5341 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 5401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 5461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 5521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 5581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 5641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 5701 aataggggtt ccgcgcacat ttcc //