LOCUS       sequence-406186-        7986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJAT plasmid with multiple cloning site for introducing custom 
            payload. One U6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7986)
  AUTHORS   Stern  D, Kim E, Behrman E
  TITLE     The Janelia Atalanta plasmids provide a simple and efficient 
            CRISPR/Cas9-mediated homology directed repair platform for 
            Drosophila
  JOURNAL   bioRxiv 2023.06.17.545412
REFERENCE   2  (bases 1 to 7986)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.06.17.545412"
COMMENT     Sequence Label: pJAT74
FEATURES             Location/Qualifiers
     source          1..7986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(943..1066)
                     /gene="mutant version of attR"
                     /label=attR6
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1158..1177)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1159..1177)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1630..2421
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2667..3255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3156..3175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3409..3426
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3458..3476
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3458..3475
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        3553..3973
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3 
                     snRNA (Port et al., 2014)"
     protein_bind    3980..4103
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4126..4131,4132..4149,4150..4156)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     promoter        4199..4303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4304..4372,4373..5164)
                     /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(4522..4541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     5685..5704
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5844..5967)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    6125..6248
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(6271..6276,6277..6294,6295..6301)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6355..7014
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(6421..6440)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        7023..7468
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(7469..7555,7556..7986,1..904)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 aaaaatccgt ttattctaca ctgatttctc cggtaaacat tacggcaaac aaacactgac
       61 aactgcacaa gttaacgtat cagcatcaga cagctctttg aacatcaacg gtgtagagga
      121 ttataaatca atctttgacg gtgacggaaa aacgtatcaa aatgtacagc agttcatcga
      181 tgaaggcaac tacagctcag gcgacaacca tacgctgaga gatcctcact acgtagaaga
      241 taaaggccac aaatacttag tatttgaagc aaacactgga actgaagatg gctaccaagg
      301 cgaagaatct ttatttaaca aagcatacta tggcaaaagc acatcattct tccgtcaaga
      361 aagtcaaaaa cttctgcaaa gcgataaaaa acgcacggct gagttagcaa acggcgctct
      421 cggtatgatt gagctaaacg atgattacac actgaaaaaa gtgatgaaac cgctgattgc
      481 atctaacaca gtaacagatg aaattgaacg cgcgaacgtc tttaaaatga acggcaaatg
      541 gtacctgttc actgactccc gcggatcaaa aatgacgatt gacggcatta cgtctaacga
      601 tatttacatg cttggttatg tttctaattc tttaactggc ccatacaagc cgctgaacaa
      661 aactggcctt gtgttaaaaa tggatcttga tcctaacgat gtaaccttta cttactcaca
      721 cttcgctgta cctcaagcga aaggaaacaa tgtcgtgatt acaagctata tgacaaacag
      781 aggattctac gcagacaaac aatcaacgtt tgcgccaagc ttcctgctga acatcaaagg
      841 caagaaaaca tctgttgtca aagacagcat ccttgaacaa ggacaattaa cagttaacaa
      901 ataaatgtca ggctccctta tacacagcca gtctgcacgt cgcatagtga ctggatatgt
      961 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     1021 tttatatcat tttacgtttc tcgttcaact ttttaataca aagttgcctc cggaacataa
     1081 tggtgcatct agatgcccat tcctgtagac ttcttaatta agacgtcaga attctcgagg
     1141 cggccgcatg tgagtctccc tatagtgagt cgtattaatt tcgcgggcgc gtcagcggga
     1201 gagccgttgt aaggcggcag actttgctca tgttaccgat gctattcgga agaacggcaa
     1261 ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg gtagcatgtt gattgtaacg
     1321 atgacagagc gttgctgcct gtgatcaatt cgggcacgaa cccagtggac ataagcctgt
     1381 tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     1441 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     1501 tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
     1561 tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
     1621 ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt
     1681 ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca
     1741 gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg
     1801 cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct
     1861 ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt
     1921 ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt
     1981 cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa
     2041 gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt
     2101 cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc
     2161 gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca
     2221 gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg
     2281 gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat
     2341 cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc
     2401 accaaggtag tcggcaaata atgtctagct agaaattcgt tcaagccgac gccgcttcgc
     2461 ggcgcggctt aactcaagcg ttagatgcac taagcacata attgctcaca gccaaactat
     2521 caggtcaagt ctgcttttat tatttttaag cgtgcataat aagccctaca caaattggga
     2581 gatatatcat gcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2641 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2701 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2761 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2821 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2881 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2941 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3001 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3061 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3121 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3181 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3241 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3301 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3361 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3421 agcgaggaaa tcgtcacctg caggaaggtt taaacgcatt taggtgacac tatagaagtg
     3481 tgtatcgctc gagggatccg aattcgaagt cttggtacgg aggggatccg atcgacaagc
     3541 cgaacatatg ggttttttgc tcacctgtga ttgctcctac tcaaatacaa aaacatcaaa
     3601 ttttctgtca ataaagcata tttatttata tttattttac aggaaagaat tccttttaaa
     3661 gtgtatttta acctataatg aaaaacgatt aaaaaaaata cataaaataa ttcgaaaatt
     3721 tttgaatagc ccaggttgat aaaaattcat ttcatacgtt ttataactta tgcccctaag
     3781 tattttttga ccatagtgtt tcaattctac attaatttta cagagtagaa tgaaacgcca
     3841 cctactcagc caagaggcga aaaggttagc tcgccaagca gagagggcgc cagtgctcac
     3901 tactttttat aattctcaac ttctttttcc agactcagtt cgtatatata gacctatttt
     3961 caatttaacg tcggggcctc aactttgtat aataaagttg aacgagaaac gtaaaatgat
     4021 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     4081 acacaacata tccagtcact atgggccgca ttaggcaccc caggctttac actttatgct
     4141 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctacg
     4201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4261 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4381 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4441 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4501 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4561 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4621 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4681 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4741 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4801 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4861 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4921 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4981 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcccgcggta tcattgcagc
     5041 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5101 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5161 gtaatggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt
     5221 gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat
     5281 gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca aggcatatat
     5341 gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt
     5401 gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa
     5461 atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac
     5521 ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttatcgatac
     5581 accgggccgt cgcatggtta ttccacttgc gtctgcgcgc ttgctctcag ataaagtctc
     5641 ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga
     5701 tatggccagt gtgccggttt ccgttatcgg ggaagaagtg gctgatctca gccaccgcga
     5761 aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataatgtc aggctccctt
     5821 atacacagcc agtctgcacg tcgcatagtg actggatatg ttgtgtttta cagtattatg
     5881 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     5941 ctcgttcaac ttttctatac aaagttgtcc agctggagcg agaccggagt cgtcttcgct
     6001 ttccgaattc gagctcggta cctcgcgaat gcatctagat atcagatctg gatcccgggc
     6061 ccgtcgactg cagaggcctg catgcaagct tgataacttg tgaagacgac catggtctca
     6121 ccatcaactt tgtatacaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     6181 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     6241 tcactatggg ccgcattagg caccccaggc tttacacttt atgcttccgg ctcgtataat
     6301 gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc taaaatggag
     6361 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     6421 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     6481 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     6541 cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga cggtgagctg
     6601 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     6661 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     6721 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     6781 tttttcgtgt cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     6841 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     6901 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     6961 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaaacgcgt
     7021 ggcacatata cctgccgttc actattattt agtgaaatga gatattatga tattttctga
     7081 attgtgatta aaaaggcaac tttatgccca tgcaacagaa actataaaaa atacagagaa
     7141 tgaaaagaaa cagatagatt ttttagttct ttaggcccgt agtctgcaaa tccttttatg
     7201 attttctatc aaacaaaaga ggaaaataga ccagttgcaa tccaaacgag agtctaatag
     7261 aatgaggtcg aaaagtaaat cgcgcgggtt tgttactgat aaagcaggca agacctaaaa
     7321 tgtgtaaagg gcaaagtgta tactttggcg tcacccctta catattttag gtcttttttt
     7381 attgtgcgta actaacttgc catcttcaaa caggagggct ggaagaagca gaccgctaac
     7441 acagtacata aaaaaggaga catgaacgat gaacatcaaa aagtttgcaa aacaagcaac
     7501 agtattaacc tttactaccg cactgctggc aggaggcgca actcaagcgt ttgcgaaaga
     7561 aacgaaccaa aagccatata aggaaacata cggcatttcc catattacac gccatgatat
     7621 gctgcaaatc cctgaacagc aaaaaaatga aaaatatcaa gttcctgaat tcgattcgtc
     7681 cacaattaaa aatatctctt ctgcaaaagg cctggacgtt tgggacagct ggccattaca
     7741 aaacgctgac ggcactgtcg caaactatca cggctaccac atcgtctttg cattagccgg
     7801 agatcctaaa aatgcggatg acacatcgat ttacatgttc tatcaaaaag tcggcgaaac
     7861 ttctattgac agctggaaaa acgctggccg cgtctttaaa gacagcgaca aattcgatgc
     7921 aaatgattct atcctaaaag accaaacaca agaatggtca ggttcagcca catttacatc
     7981 tgacgg
//
