LOCUS       sequence-406178-        7560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJAT plasmid with multiple cloning site for introducing custom 
            payload. No U6 promoter. Restriction sites present to introduce 
            flanking U6 promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7560)
  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 7560)
  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: pJAT77
FEATURES             Location/Qualifiers
     source          1..7560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     569..588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(807..911)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(join(954..960,961..978,979..984))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1007..1130)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1208..1226)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1209..1226)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1258..1275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1429..2017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1509..1528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2263..3054)
                     /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"
     primer_bind     3507..3526
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3507..3525
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3618..3741
                     /gene="mutant version of attR"
                     /label=attR6
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(join(3780..5114,5115..5201))
                     /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"
     promoter        complement(5202..5647)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             complement(5656..6315)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6230..6249
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6369..6375,6376..6393,6394..6399))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(6422..6545)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    6703..6826
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6966..6985)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(join(7506..7560,1..737,738..806))
                     /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"
ORIGIN
        1 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
       61 ggccccagtg ctgcaatgat accgcgggac ccacgctcac cggctccaga tttatcagca
      121 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
      181 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
      241 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
      301 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
      361 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
      421 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
      481 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
      541 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
      601 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
      661 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
      721 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
      781 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
      841 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
      901 ggggttccgc gtagctcctg aaaatctcga cggatcctaa ctcaaaatcc acacattata
      961 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gcggcccata gtgactggat
     1021 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     1081 gatatttata tcattttacg tttctcgttc aactttatta tacaaagttg aggcctatgt
     1141 tcggcttgtc gatcggatcc cctccgtacc aagacttcga attcggatcc ctcgagcgat
     1201 acacacttct atagtgtcac ctaaatgcgt ttaaaccttc ctgcaggtga cgatttcctc
     1261 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1321 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1381 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1441 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1501 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1561 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1621 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1681 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1741 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1801 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1861 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1921 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     1981 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2041 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgcatgatat
     2101 atctcccaat ttgtgtaggg cttattatgc acgcttaaaa ataataaaag cagacttgac
     2161 ctgatagttt ggctgtgagc aattatgtgc ttagtgcatc taacgcttga gttaagccgc
     2221 gccgcgaagc ggcgtcggct tgaacgaatt tctagctaga cattatttgc cgactacctt
     2281 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
     2341 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
     2401 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     2461 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     2521 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     2581 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     2641 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agatacctgc
     2701 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     2761 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcgctctc
     2821 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     2881 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggcttca ggccgccatc
     2941 cactgcggag ccgtacaaat gtacggccag caacgtcggt tcgagatggc gctcgatgac
     3001 gccaactacc tctgatagtt gagtcgatac ttcggcgatc accgcttccc ccatgatgtt
     3061 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa
     3121 acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacccc
     3181 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
     3241 ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca gcttacgaac
     3301 cgaacaggct tatgtccact gggttcgtgc ccgaattgat cacaggcagc aacgctctgt
     3361 catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct
     3421 tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg
     3481 ctctcccgct gacgcgcccg cgaaattaat acgactcact atagggagac tcacatgcgg
     3541 ccgcctcgag aattctgacg tcttaattaa gaagtctaca ggaatgggca tctagatgca
     3601 ccattatgtt ccggaggcaa ctttgtatta aaaagttgaa cgagaaacgt aaaatgatat
     3661 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     3721 acaacatatc cagtcactat gcgacgtgca gactggctgt gtataaggga gcctgacatt
     3781 tatttgttaa ctgttaattg tccttgttca aggatgctgt ctttgacaac agatgttttc
     3841 ttgcctttga tgttcagcag gaagcttggc gcaaacgttg attgtttgtc tgcgtagaat
     3901 cctctgtttg tcatatagct tgtaatcacg acattgtttc ctttcgcttg aggtacagcg
     3961 aagtgtgagt aagtaaaggt tacatcgtta ggatcaagat ccatttttaa cacaaggcca
     4021 gttttgttca gcggcttgta tgggccagtt aaagaattag aaacataacc aagcatgtaa
     4081 atatcgttag acgtaatgcc gtcaatcgtc atttttgatc cgcgggagtc agtgaacagg
     4141 taccatttgc cgttcatttt aaagacgttc gcgcgttcaa tttcatctgt tactgtgtta
     4201 gatgcaatca gcggtttcat cacttttttc agtgtgtaat catcgtttag ctcaatcata
     4261 ccgagagcgc cgtttgctaa ctcagccgtg cgttttttat cgctttgcag aagtttttga
     4321 ctttcttgac ggaagaatga tgtgcttttg ccatagtatg ctttgttaaa taaagattct
     4381 tcgccttggt agccatcttc agttccagtg tttgcttcaa atactaagta tttgtggcct
     4441 ttatcttcta cgtagtgagg atctctcagc gtatggttgt cgcctgagct gtagttgcct
     4501 tcatcgatga actgctgtac attttgatac gtttttccgt caccgtcaaa gattgattta
     4561 taatcctcta caccgttgat gttcaaagag ctgtctgatg ctgatacgtt aacttgtgca
     4621 gttgtcagtg tttgtttgcc gtaatgttta ccggagaaat cagtgtagaa taaacggatt
     4681 tttccgtcag atgtaaatgt ggctgaacct gaccattctt gtgtttggtc ttttaggata
     4741 gaatcatttg catcgaattt gtcgctgtct ttaaagacgc ggccagcgtt tttccagctg
     4801 tcaatagaag tttcgccgac tttttgatag aacatgtaaa tcgatgtgtc atccgcattt
     4861 ttaggatctc cggctaatgc aaagacgatg tggtagccgt gatagtttgc gacagtgccg
     4921 tcagcgtttt gtaatggcca gctgtcccaa acgtccaggc cttttgcaga agagatattt
     4981 ttaattgtgg acgaatcgaa ttcaggaact tgatattttt catttttttg ctgttcaggg
     5041 atttgcagca tatcatggcg tgtaatatgg gaaatgccgt atgtttcctt atatggcttt
     5101 tggttcgttt ctttcgcaaa cgcttgagtt gcgcctcctg ccagcagtgc ggtagtaaag
     5161 gttaatactg ttgcttgttt tgcaaacttt ttgatgttca tcgttcatgt ctcctttttt
     5221 atgtactgtg ttagcggtct gcttcttcca gccctcctgt ttgaagatgg caagttagtt
     5281 acgcacaata aaaaaagacc taaaatatgt aaggggtgac gccaaagtat acactttgcc
     5341 ctttacacat tttaggtctt gcctgcttta tcagtaacaa acccgcgcga tttacttttc
     5401 gacctcattc tattagactc tcgtttggat tgcaactggt ctattttcct cttttgtttg
     5461 atagaaaatc ataaaaggat ttgcagacta cgggcctaaa gaactaaaaa atctatctgt
     5521 ttcttttcat tctctgtatt ttttatagtt tctgttgcat gggcataaag ttgccttttt
     5581 aatcacaatt cagaaaatat cataatatct catttcacta aataatagtg aacggcaggt
     5641 atatgtgcca cgcgtttacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     5701 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     5761 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     5821 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     5881 cacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     5941 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     6001 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     6061 ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag
     6121 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     6181 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     6241 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     6301 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgacggat cctaactcaa
     6361 aatccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgcggc
     6421 ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat
     6481 ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt ttgtatacaa
     6541 agttgatggt gagaccatgg tcgtcttcac aagttatcaa gcttgcatgc aggcctctgc
     6601 agtcgacggg cccgggatcc agatctgata tctagatgca ttcgcgaggt accgagctcg
     6661 aattcggaaa gcgaagacga ctccggtctc gctccagctg gacaactttg tatagaaaag
     6721 ttgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa
     6781 aacagactac ataatactgt aaaacacaac atatccagtc actatgcgac gtgcagactg
     6841 gctgtgtata agggagcctg acattatatt ccccagaaca tcaggttaat ggcgtttttg
     6901 atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga aaccggcaca
     6961 ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac caccgggtaa
     7021 agttcacggg agactttatc tgagagcaag cgcgcagacg caagtggaat aaccatgcga
     7081 cggcccggtg tatcgataat atcactctgt acatccacaa acagacgata acggctctct
     7141 cttttatagg tgtaaacctt aaactgcatt tcaccagccc ctgttctcgt cagcaaaaga
     7201 gccgttcatt tcaataaacc gggcgacctc agccatccct tcctgatttt ccgctttcca
     7261 gcgttcggca cgcagacgac gggcttcatt ctgcatggtt gtgcttacca gaccggagat
     7321 attgacatca tatatgcctt gagcaactga tagctgtcgc tgtcaactgt cactgtaata
     7381 cgctgcttca tagcatacct ctttttgaca tacttcgggt atacatatca gtatatattc
     7441 ttataccgca aaaatcagcg cgcaaatacg catactgtta tctggctttt agtaagccgg
     7501 atccattacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
//
