LOCUS       sequence-367524-        4029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty donor plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4029)
  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 4029)
  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: puc19_N_tag
FEATURES             Location/Qualifiers
     source          1..4029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..32)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        100..204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(205..273,274..1065)
                     /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(423..442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1236..1824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1725..1744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1978..1995
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2112..2133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2148..2153,2154..2171,2172..2178)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2186..2202
                     /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     2191..2213
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2210..2226
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2210..2226
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2264..2398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2288..2307)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2342..2361
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2399..2998)
                     /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(2482..2502)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2979..2998
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             3353..3418
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3455..3544
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(3639..3656)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3639..3655)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3648..3670)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3864..3883)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3983..4005
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
       61 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      421 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
      481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
      601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
      661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
      721 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
      781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
      841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
      901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
      961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1381 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     1981 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     2041 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     2101 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     2161 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     2221 tatgaccatg attacgccag aagacagaaa cacccaggta agtgatccag acatgataag
     2281 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     2341 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttct
     2401 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     2461 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca
     2521 ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc
     2581 ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct
     2641 ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac
     2701 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
     2761 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
     2821 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt
     2881 ccgggtcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
     2941 cggcccgggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcattt
     3001 tgattgtctc tggattagac gactgctggc tgatggagcg gctttgtgtc gggaggagta
     3061 tccacacagc acaagaactc aaacggtagt gatatgaaaa ctggtcccgc agcccgtttt
     3121 ataaagcccg cttagcgcta aaaccttatc agccgcaatg ctctccgctc tcaagtcgcg
     3181 ttcaaaactt ttaccatata tgggtatgaa ttgttcgccg attgggtttt catttacgga
     3241 gaatttcctc cgcaactggg tgttcgtgct tgtgtgtgtc ttgtatatcg ccatgaagag
     3301 tgtacctaat tttaatcaac cgatttctct ctcttccgca ggaatcaaaa tggactacaa
     3361 agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg atgacaaggg
     3421 tggatctgga ggttccggcg gctcaggggg tagttaccca tacgatgttc ctgactatgc
     3481 gggctatccc tatgacgtcc cggactatgc aggatcctat ccatatgacg ttccagatta
     3541 cgcttccggt ggctctggct cttcaggctc atattattat gtcttctgca tgcctgcagg
     3601 tcgactctag aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt
     3661 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3721 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3781 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3841 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3901 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3961 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4021 ccgaaacgc
//
