LOCUS       sequence-369153-        5192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid to endogenously tag Shu gene in Drosophila 
            melanogaster ovarian somatic sheet (OSS) cells. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5192)
  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 5192)
  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: "Nucleic 
            Acids Res. 2022 May 27. pii: 6594084. doi: 10.1093/nar/gkac448."
COMMENT     Sequence Label: Donor_N_FH_Shu
FEATURES             Location/Qualifiers
     source          1..5192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        join(541..546,547..564,565..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    1194..1328
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1218..1237)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1272..1291
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1329..1928)
                     /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(1412..1432)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1909..1928
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             2283..2348
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2385..2474
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(3195..3212)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3195..3211)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3204..3226)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3420..3439)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3539..3561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3599..3617)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3685..3789
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3790..3858,3859..4650)
                     /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(4008..4027)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4821..5192,1..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 gtgaacaagc
      661 gagcattcgc atactattgt attgatcatt gcttagaata gccctccctt aatatacaaa
      721 tcttagccat accatatcgc acggtgagca cttcacaagg ccattaagaa tgcattcctc
      781 attatgtttc ctagtttttt attgttacac acaccaaaac aattctctca atggagtgct
      841 cgtagttgga gagcgataag tgttcagttt taaataaaga ttcatataaa aagcaagtat
      901 atctggccta gtgtttcccc gtctggattg gaatacatta tacattttcg gaagatatct
      961 cgagttgcag tgctcgcgcc ctaataaaga tatatagttc tttgtttgaa ttcgataact
     1021 tccggacaac agcgaccaat cgattcgcta aagctatcaa gcaatcgaat tccagttgat
     1081 accatcgctg gaaagctctt ttctcaaacg catccaaaag aatagcgcca actcagcgca
     1141 ttccggaatt cgtttcgtaa atattctgta aattttgcga tacccaggta agtgatccag
     1201 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1261 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     1321 aacaagttct aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc
     1381 tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg
     1441 gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg
     1501 acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac
     1561 cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc
     1621 agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct
     1681 tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc
     1741 ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg
     1801 atgtggcggt ccgggtcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg
     1861 agggtgcgta cggcccgggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac
     1921 tcggtcattt tgattgtctc tggattagac gactgctggc tgatggagcg gctttgtgtc
     1981 gggaggagta tccacacagc acaagaactc aaacggtagt gatatgaaaa ctggtcccgc
     2041 agcccgtttt ataaagcccg cttagcgcta aaaccttatc agccgcaatg ctctccgctc
     2101 tcaagtcgcg ttcaaaactt ttaccatata tgggtatgaa ttgttcgccg attgggtttt
     2161 catttacgga gaatttcctc cgcaactggg tgttcgtgct tgtgtgtgtc ttgtatatcg
     2221 ccatgaagag tgtacctaat tttaatcaac cgatttctct ctcttccgca ggaatcaaaa
     2281 tggactacaa agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg
     2341 atgacaaggg tggatctgga ggttccggcg gctcaggggg tagttaccca tacgatgttc
     2401 ctgactatgc gggctatccc tatgacgtcc cggactatgc aggatcctat ccatatgacg
     2461 ttccagatta cgcttccggt ggctctggct cttcaggctc aatggaagaa aacttcgaac
     2521 cgtacacccc acagttgctc aagaatccct tgtcctattc gtgagtaata agatacatat
     2581 taaaggcact catgaccacg tgcaactcat tctgcaggga tctcgtgaaa aagggcgttg
     2641 agttcgaggt cgacaattcg cagcagaacc atgccaggga tctgggtctc gacagcgaca
     2701 gcgacagcga ctacgaggat gcgctggacg tggacggcga ggagctacgt tcaccgtgga
     2761 catattcatt cgacgaactg cgtgcgctga tgagcgagat cgacgaaaac atttacaagc
     2821 ggatcactcg caccggtcac gtggatcggg aggcggtgcc gaacaaggct agggtctcgg
     2881 ttcggtacag tggctactgg gagggcgaga ccgctccctt cgactcctcc ctgctgcggg
     2941 gcagcaaatt cgtgtttgaa actggccagg gaacggtggt tgaaggcctg gaggtcgccg
     3001 tccgcagcat gcgtccttac gagcaggccg agtttataat ctcctacaag ctgcttttcg
     3061 gagaactcgg ttgccctccg cggatcaagc ccaaggcgga tgcgctcttt aaagtggagg
     3121 taattgacta ctcgctcata ggagatgcga agggcatcga tgctatagga tccccgggta
     3181 ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3241 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3301 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     3361 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     3421 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3481 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3541 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3601 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3661 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3721 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3781 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3841 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3901 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3961 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4021 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4081 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4141 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4201 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4261 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4321 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4381 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4441 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4501 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4561 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4621 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4681 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4741 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4801 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4861 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4921 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4981 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5041 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5101 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5161 cccagcttgg agcgaacgac ctacaccgaa ct
//
