LOCUS       sequence-406380-        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"
     CDS             complement(join(151..942,943..1011))
                     /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     774..793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1012..1116)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(join(1159..1165,1166..1183,1184..1189))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1212..1335)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1413..1431)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1414..1431)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1463..1480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1634..2222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1714..1733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2468..3259)
                     /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     3712..3731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3712..3730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3823..3946
                     /gene="mutant version of attR"
                     /label=attR6
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(join(3985..5319,5320..5406))
                     /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(5407..5852)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             complement(5861..6520)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6435..6454
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6574..6580,6581..6598,6599..6604))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(6627..6750)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    6908..7031
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7171..7190)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt
       61 cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac
      121 tgttatctgg cttttagtaa gccggatcca ttaccaatgc ttaatcagtg aggcacctat
      181 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
      241 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gggacccacg
      301 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
      361 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
      421 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      481 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      541 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      601 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      661 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      721 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      781 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      841 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      901 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      961 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1081 atgtatttag aaaaataaac aaataggggt tccgcgtagc tcctgaaaat ctcgacggat
     1141 cctaactcaa aatccacaca ttatacgagc cggaagcata aagtgtaaag cctggggtgc
     1201 ctaatgcggc ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt
     1261 tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcaactt
     1321 tattatacaa agttgaggcc tatgttcggc ttgtcgatcg gatcccctcc gtaccaagac
     1381 ttcgaattcg gatccctcga gcgatacaca cttctatagt gtcacctaaa tgcgtttaaa
     1441 ccttcctgca ggtgacgatt tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1501 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1561 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2041 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     2101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2281 aagggatttt ggtcatgcat gatatatctc ccaatttgtg tagggcttat tatgcacgct
     2341 taaaaataat aaaagcagac ttgacctgat agtttggctg tgagcaatta tgtgcttagt
     2401 gcatctaacg cttgagttaa gccgcgccgc gaagcggcgt cggcttgaac gaatttctag
     2461 ctagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt
     2521 ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag
     2581 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
     2641 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
     2701 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
     2761 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
     2821 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
     2881 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
     2941 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
     3001 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
     3061 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
     3121 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
     3181 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
     3241 cgatcaccgc ttcccccatg atgtttaact ttgttttagg gcgactgccc tgctgcgtaa
     3301 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     3361 atgcccgagg catagactgt accccaaaaa aacagtcata acaagccatg aaaaccgcca
     3421 ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac
     3481 gctacttgca ttacagctta cgaaccgaac aggcttatgt ccactgggtt cgtgcccgaa
     3541 ttgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc tccgcgagat
     3601 catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat cggtaacatg
     3661 agcaaagtct gccgccttac aacggctctc ccgctgacgc gcccgcgaaa ttaatacgac
     3721 tcactatagg gagactcaca tgcggccgcc tcgagaattc tgacgtctta attaagaagt
     3781 ctacaggaat gggcatctag atgcaccatt atgttccgga ggcaactttg tattaaaaag
     3841 ttgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa
     3901 aacagactac ataatactgt aaaacacaac atatccagtc actatgcgac gtgcagactg
     3961 gctgtgtata agggagcctg acatttattt gttaactgtt aattgtcctt gttcaaggat
     4021 gctgtctttg acaacagatg ttttcttgcc tttgatgttc agcaggaagc ttggcgcaaa
     4081 cgttgattgt ttgtctgcgt agaatcctct gtttgtcata tagcttgtaa tcacgacatt
     4141 gtttcctttc gcttgaggta cagcgaagtg tgagtaagta aaggttacat cgttaggatc
     4201 aagatccatt tttaacacaa ggccagtttt gttcagcggc ttgtatgggc cagttaaaga
     4261 attagaaaca taaccaagca tgtaaatatc gttagacgta atgccgtcaa tcgtcatttt
     4321 tgatccgcgg gagtcagtga acaggtacca tttgccgttc attttaaaga cgttcgcgcg
     4381 ttcaatttca tctgttactg tgttagatgc aatcagcggt ttcatcactt ttttcagtgt
     4441 gtaatcatcg tttagctcaa tcataccgag agcgccgttt gctaactcag ccgtgcgttt
     4501 tttatcgctt tgcagaagtt tttgactttc ttgacggaag aatgatgtgc ttttgccata
     4561 gtatgctttg ttaaataaag attcttcgcc ttggtagcca tcttcagttc cagtgtttgc
     4621 ttcaaatact aagtatttgt ggcctttatc ttctacgtag tgaggatctc tcagcgtatg
     4681 gttgtcgcct gagctgtagt tgccttcatc gatgaactgc tgtacatttt gatacgtttt
     4741 tccgtcaccg tcaaagattg atttataatc ctctacaccg ttgatgttca aagagctgtc
     4801 tgatgctgat acgttaactt gtgcagttgt cagtgtttgt ttgccgtaat gtttaccgga
     4861 gaaatcagtg tagaataaac ggatttttcc gtcagatgta aatgtggctg aacctgacca
     4921 ttcttgtgtt tggtctttta ggatagaatc atttgcatcg aatttgtcgc tgtctttaaa
     4981 gacgcggcca gcgtttttcc agctgtcaat agaagtttcg ccgacttttt gatagaacat
     5041 gtaaatcgat gtgtcatccg catttttagg atctccggct aatgcaaaga cgatgtggta
     5101 gccgtgatag tttgcgacag tgccgtcagc gttttgtaat ggccagctgt cccaaacgtc
     5161 caggcctttt gcagaagaga tatttttaat tgtggacgaa tcgaattcag gaacttgata
     5221 tttttcattt ttttgctgtt cagggatttg cagcatatca tggcgtgtaa tatgggaaat
     5281 gccgtatgtt tccttatatg gcttttggtt cgtttctttc gcaaacgctt gagttgcgcc
     5341 tcctgccagc agtgcggtag taaaggttaa tactgttgct tgttttgcaa actttttgat
     5401 gttcatcgtt catgtctcct tttttatgta ctgtgttagc ggtctgcttc ttccagccct
     5461 cctgtttgaa gatggcaagt tagttacgca caataaaaaa agacctaaaa tatgtaaggg
     5521 gtgacgccaa agtatacact ttgcccttta cacattttag gtcttgcctg ctttatcagt
     5581 aacaaacccg cgcgatttac ttttcgacct cattctatta gactctcgtt tggattgcaa
     5641 ctggtctatt ttcctctttt gtttgataga aaatcataaa aggatttgca gactacgggc
     5701 ctaaagaact aaaaaatcta tctgtttctt ttcattctct gtatttttta tagtttctgt
     5761 tgcatgggca taaagttgcc tttttaatca caattcagaa aatatcataa tatctcattt
     5821 cactaaataa tagtgaacgg caggtatatg tgccacgcgt ttacgccccg ccctgccact
     5881 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     5941 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     6001 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     6061 tgaaactcac ccagggattg gctgacacga aaaacatatt ctcaataaac cctttaggga
     6121 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     6181 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     6241 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     6301 ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     6361 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     6421 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     6481 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     6541 aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa gcataaagtg
     6601 taaagcctgg ggtgcctaat gcggcccata gtgactggat atgttgtgtt ttacagtatt
     6661 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
     6721 tttctcgttc aacttttgta tacaaagttg atggtgagac catggtcgtc ttcacaagtt
     6781 atcaagcttg catgcaggcc tctgcagtcg acgggcccgg gatccagatc tgatatctag
     6841 atgcattcgc gaggtaccga gctcgaattc ggaaagcgaa gacgactccg gtctcgctcc
     6901 agctggacaa ctttgtatag aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat
     6961 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
     7021 cagtcactat gcgacgtgca gactggctgt gtataaggga gcctgacatt atattcccca
     7081 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc
     7141 ttccccgata acggaaaccg gcacactggc catatcggtg gtcatcatgc gccagctttc
     7201 atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaga gcaagcgcgc
     7261 agacgcaagt ggaataacca tgcgacggcc cggtgtatcg ataatatcac tctgtacatc
     7321 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc
     7381 agcccctgtt ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca
     7441 tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca
     7501 tggttgtgct taccagaccg gagatattga catcatatat gccttgagca actgatagct
//
