LOCUS       sequence-367687-        4575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid to endogenously tag VIR 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 4575)
  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 4575)
  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_C_FH_VIR
FEATURES             Location/Qualifiers
     source          1..4575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             744..809
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             828..854
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             864..920
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             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(921..940)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1417..1437
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2161..2178)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2161..2177)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2170..2192)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2386..2405)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2505..2527
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2565..2583)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2651..2755
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2756..2824,2825..3616)
                     /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(2974..2993)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3787..4375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4276..4295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4529..4546
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc aggagaagat
      241 gtttgtgtcg tcggtaaggg gtcgtggttt tgctcggcca ccaccttcca ggggcgattt
      301 gttccgctcc cggccgccca acacctcgcg accgccatcg ctgcacgtgg acgacttcct
      361 ggccttggaa acttgtggcg cccagcccac aggacccacc ggctacaaca agataccctc
      421 aatgctgaga ggctcccggg tgggtcgcaa ccgaggatct cgaatctcgg cagcagctgc
      481 ctttaggtaa agtgtcacga ttaccactta gcaggataac gctaatgggc tctttttact
      541 cttttaacag gcaaaagaag atgatgcgca ttggttcgcc ctccagctgg gcagagtccc
      601 ccggatcgta tcggtctgcc tcggactccc acttcagtag cagcgactcg cactattcgt
      661 cgccccacta cagtggacgg ccgcgtggac gaggcctgcg ttccaggcca tcctacctga
      721 gaaccagcgg aggaggttcc ggtgactaca aggaccacga cggcgattat aaggatcacg
      781 acatcgacta caaagacgac gatgacaagg ctggcgccag cagcgcctac ccatacgacg
      841 taccagatta cgctgctgga tctgccacga acttctctct gttaaagcaa gcaggagacg
      901 tggaagaaaa ccccggtcct atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
      961 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     1021 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     1081 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     1141 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     1201 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     1261 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     1321 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     1381 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     1441 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     1501 cccgcaagcc cggtgcctga tagaggacat tgcagcccaa cccgcttctc attgacactt
     1561 cccgtaaact cggcaaacga accagaacta gatcctaaga tgcccgctgg gcagctgaat
     1621 tccattactt aataacgtgt gttgctttcg tattagttcg ctttacttcc cactttgtat
     1681 gtaagccaaa agttagtttt ccgtaattac actgtgcaaa cggacgtcgt gcgactccat
     1741 gggcagctcc tcgttgctga ctatctgctc gttgagcgac aacgccatca gcgagatctt
     1801 cttgtgcgga tacttctccg cgtgctgctt gaggaacttg tcgtagtagc ccatgccatg
     1861 gcccatccga gctccgcagc gggtgaaggc tacaccgggc acaatgaaga gatcgatgcc
     1921 gtgccctgtg gtgatcgagt gtgtgtgtgt gtgggtttag aaggagctct gatgtatgcc
     1981 tagccactca ccgttggtca tggcatcctc gcgtgcctcc ttgaagtccg gctgcttgat
     2041 gttccacttg gtgagaggca ggctgtcgta ctcttccatg ccgcgcagcc gcaccatctt
     2101 catcctgctg ccctcgtagg tgggcacaaa gaaggatccc cgggtaccga gctcgaattc
     2161 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2221 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2281 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     2341 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     2401 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2461 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2521 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     2581 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2641 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2701 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2761 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2821 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2881 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2941 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3001 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3061 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3121 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3181 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3241 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3301 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3361 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3421 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3481 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3541 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3601 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3661 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3721 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4441 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4501 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4561 cccaatacgc aaacc
//
