LOCUS       sequence-367690-        4983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid to endogenously tag Nbr gene in Drosophila 
            melanogaster at the C-terminal. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4983)
  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 4983)
  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_Nbr
FEATURES             Location/Qualifiers
     source          1..4983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(73..90)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(73..89)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(82..104)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(298..317)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     417..439
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(477..495)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        563..667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(668..736,737..1528)
                     /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(886..905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1699..2287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2188..2207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2441..2458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2575..2596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2611..2616,2617..2634,2635..2641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2649..2665
                     /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     2654..2676
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2673..2689
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2673..2689
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3474..3539
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3558..3584
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3594..3650
                     /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             3651..4250
                     /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(3651..3670)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4147..4167
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ttggatgcca agtcctgcag tgcatgcctg caggtcgact ctagaggatc cccgggtacc
       61 gagctcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      121 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      181 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      241 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
      301 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      361 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      421 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
      481 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
      541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1381 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1681 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1801 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2101 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2281 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2341 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2401 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2461 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2521 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2581 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2641 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     2701 cagtagatgt gatatgtacc tgaccatgga cctgcccgac gagtgcctta tcattgtgaa
     2761 caaggctgat gagttcgatc gtatgttata ccatctgcag caggagtgcg tcatatacct
     2821 ggattctgaa tggatgcaga gcgtgtgcgg agacaatcag ctgtgcgtgc tacagatcgc
     2881 caccggccac aatgtctacc tgatcgattg cctggcgcgg gagagcttac gctcggaaca
     2941 ctggcgcttg ctgggcgcga atatctttaa taacgtgaac atccgtaagg tggggttctc
     3001 tatggtcagt gatctcagtg tattgcagcg gtcactgccc ctgcaactgc gccttcaaat
     3061 gcctcaccac tacttggacc ttcgcaatct ctggttggag ttaaaaaagc agcgcttcgg
     3121 cgttgaactt cccttcggca atgttaaccg ggcaggagac gcccttacgg atctctcatt
     3181 ggcatgcctt ggcaagaaac tgaacaaatc aaaccagtgc tcaaactggg ccaatagacc
     3241 gctgcgtcgc gagcagattc tttatgcagg tgagtcaggg ataggccttt tgattgaatg
     3301 tagcaagact aattttctac ttaacagcca tcgatgcgcg gtgtttgatg ctgatctaca
     3361 acacgctcat tgaacgcgtg tccttcatac aagcggttat cgaaaagagc atcgccagca
     3421 ataactttct caggcggggt gcccatgtta agaccagcgg aggaggttcc ggtgactaca
     3481 aggaccacga cggcgattat aaggatcacg acatcgacta caaagacgac gatgacaagg
     3541 ctggcgccag cagcgcctac ccatacgacg taccagatta cgctgctgga tctgccacga
     3601 acttctctct gttaaagcaa gcaggagacg tggaagaaaa ccccggtcct atgaccgagt
     3661 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     3721 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     3781 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     3841 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     3901 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     3961 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4021 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4081 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4141 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4201 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga tatttgagga
     4261 gctggcctct gttataccca tgctcctttt gatggatagc gattatcacg aagacgacaa
     4321 aggcgttcgc ccaacattta ttaagtgcaa atgttatatg tagttgcaaa gagtatttaa
     4381 catatattta gtgactagcc aggcagctgg gttagtgccg agttaggcag tttgtacaca
     4441 tgttacaaat cagaaggaaa ctatgtgact gcgagcagga tatgaaagaa tgaattttat
     4501 tacgttatga tttcaagctc actgtatctc catactttaa atacttgcat tattccaatt
     4561 cgtaaattcg cgtattattt gaattcacgc atgatcaaaa ctgaataaaa gcaatgaata
     4621 ggattgaaca attgtcggga gctaaaatag ttaaaataac attggcatcg gaataatcac
     4681 attacaacgt tgtgcgatta taccagcggt gtggcgtact tgctattcca gtcccgctcg
     4741 aaaacattcc gcagatcgct tcttaaggta ttagtggtct cggtctcgaa ggtttcgctt
     4801 agcaccagtc caatgccagc ggtgtccgtg aagtagtctc cactccagtt agaagttccg
     4861 atgtaagcca ctcgatctgt aaccatgtac ttgttgtggt tgacacggcc aaagggaatc
     4921 ttctcctggc tcgaatcagt cggcactatg aatcgacgaa tttgtatatc aattttgtct
     4981 tct
//
