LOCUS       sequence-367684-        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"
     protein_bind    97..118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(133..138,139..156,157..163)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    171..187
                     /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     176..198
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     195..211
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     195..211
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    786..920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(810..829)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     864..883
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(921..1520)
                     /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(1004..1024)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1501..1520
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             1875..1940
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1977..2066
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(2787..2804)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2787..2803)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2796..2818)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3012..3031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3131..3153
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3191..3209)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3277..3381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3382..3450,3451..4242)
                     /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(3600..3619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4413..5001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4902..4921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5155..5172
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
       61 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      121 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      181 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
      241 gggtgaacaa gcgagcattc gcatactatt gtattgatca ttgcttagaa tagccctccc
      301 ttaatataca aatcttagcc ataccatatc gcacggtgag cacttcacaa ggccattaag
      361 aatgcattcc tcattatgtt tcctagtttt ttattgttac acacaccaaa acaattctct
      421 caatggagtg ctcgtagttg gagagcgata agtgttcagt tttaaataaa gattcatata
      481 aaaagcaagt atatctggcc tagtgtttcc ccgtctggat tggaatacat tatacatttt
      541 cggaagatat ctcgagttgc agtgctcgcg ccctaataaa gatatatagt tctttgtttg
      601 aattcgataa cttccggaca acagcgacca atcgattcgc taaagctatc aagcaatcga
      661 attccagttg ataccatcgc tggaaagctc ttttctcaaa cgcatccaaa agaatagcgc
      721 caactcagcg cattccggaa ttcgtttcgt aaatattctg taaattttgc gatacccagg
      781 taagtgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
      841 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
      901 taagctgcaa taaacaagtt ctaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
      961 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     1021 ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     1081 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg
     1141 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     1201 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     1261 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     1321 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     1381 gtgacccgct cgatgtggcg gtccgggtcg acggtgtggc gcgtggcggg gtagtcggcg
     1441 aacgcggcgg cgagggtgcg tacggcccgg gggacgtcgt cgcgggtggc gaggcgcacc
     1501 gtgggcttgt actcggtcat tttgattgtc tctggattag acgactgctg gctgatggag
     1561 cggctttgtg tcgggaggag tatccacaca gcacaagaac tcaaacggta gtgatatgaa
     1621 aactggtccc gcagcccgtt ttataaagcc cgcttagcgc taaaacctta tcagccgcaa
     1681 tgctctccgc tctcaagtcg cgttcaaaac ttttaccata tatgggtatg aattgttcgc
     1741 cgattgggtt ttcatttacg gagaatttcc tccgcaactg ggtgttcgtg cttgtgtgtg
     1801 tcttgtatat cgccatgaag agtgtaccta attttaatca accgatttct ctctcttccg
     1861 caggaatcaa aatggactac aaagaccatg acggtgatta taaagatcat gacatcgatt
     1921 acaaggatga cgatgacaag ggtggatctg gaggttccgg cggctcaggg ggtagttacc
     1981 catacgatgt tcctgactat gcgggctatc cctatgacgt cccggactat gcaggatcct
     2041 atccatatga cgttccagat tacgcttccg gtggctctgg ctcttcaggc tcaatggaag
     2101 aaaacttcga accgtacacc ccacagttgc tcaagaatcc cttgtcctat tcgtgagtaa
     2161 taagatacat attaaaggca ctcatgacca cgtgcaactc attctgcagg gatctcgtga
     2221 aaaagggcgt tgagttcgag gtcgacaatt cgcagcagaa ccatgccagg gatctgggtc
     2281 tcgacagcga cagcgacagc gactacgagg atgcgctgga cgtggacggc gaggagctac
     2341 gttcaccgtg gacatattca ttcgacgaac tgcgtgcgct gatgagcgag atcgacgaaa
     2401 acatttacaa gcggatcact cgcaccggtc acgtggatcg ggaggcggtg ccgaacaagg
     2461 ctagggtctc ggttcggtac agtggctact gggagggcga gaccgctccc ttcgactcct
     2521 ccctgctgcg gggcagcaaa ttcgtgtttg aaactggcca gggaacggtg gttgaaggcc
     2581 tggaggtcgc cgtccgcagc atgcgtcctt acgagcaggc cgagtttata atctcctaca
     2641 agctgctttt cggagaactc ggttgccctc cgcggatcaa gcccaaggcg gatgcgctct
     2701 ttaaagtgga ggtaattgac tactcgctca taggagatgc gaagggcatc gatgctatag
     2761 gatccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2821 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2881 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2941 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3001 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     3061 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3121 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3181 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3241 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3301 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3361 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3421 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3481 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3541 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3601 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3661 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3721 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3781 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3841 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3901 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3961 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4021 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4081 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4141 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4201 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4261 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4321 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4381 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4441 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4501 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4561 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4621 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4681 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4741 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4801 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4861 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4921 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4981 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5041 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5101 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5161 tcagtgagcg aggaagcgga agagcgccca at
//
