LOCUS       sequence-135059-        4341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gap-repair recombineering for Drosophila transgenesis, contains loxP
            cassette with GFP, Amp resistant, with 5xUAS and HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4341)
  AUTHORS   Haussmann IU, Ustaoglu P, Brauer U, Hemani Y, Dix TC, Soller M
  TITLE     Plasmid-based gap-repair recombineered transgenes reveal a central 
            role for introns in mutually exclusive alternative splicing in Down 
            Syndrome Cell Adhesion Molecule exon 4.
  JOURNAL   Nucleic Acids Res. 2018 Dec 12. pii: 5239037. doi: 
            10.1093/nar/gky1254.
  PUBMED    30541104
REFERENCE   2  (bases 1 to 4341)
  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. 2018 Dec 12. pii: 5239037. doi: 10.1093/nar/gky1254."
COMMENT     Sequence Label: pUC 3GLA UAS HAi
FEATURES             Location/Qualifiers
     source          1..4341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    188..257
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(358..377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     477..499
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(537..555)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        623..727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(728..796,797..1588)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(946..965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1759..2347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2248..2267
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2404..2498
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    2423..2517
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        2536..2774
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     2721..2742
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             2851..2877
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2887..2913
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2893..2919
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     protein_bind    3149..3182
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3418..3427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3439..4149
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             3439..4149
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(3478..3499)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3739..3758)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4086..4107
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    4296..4329
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 tgcccgccgt gaccgtcgag aacccgctga cgctgccccg cgtatccgca cccgccgacg
       61 ccgtcgcacg tcccgtgctc accgtgacca ccgcgcccag cggtttcgag ggcgagggct
      121 tcccggtgcg ccgcgcgttc gccgggatca actaccgcca cctcgacccg ttcatcatga
      181 tggaccagat gggtgaggtg gagtacgcgc ccggggagcc caagggcacg ccctggcacc
      241 cgcaccgcgg cttcgagacc tgacctacat cgtaccggct tgacgacctt ggagcgactg
      301 tcgttagttc cgcgcgattc ggtgcggtat ttcacaccgc atatggtgca ctctcagtac
      361 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      421 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      481 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
      541 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
      601 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      661 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      721 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      781 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      841 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      901 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      961 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1021 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1081 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1141 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1201 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1261 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1321 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1381 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1441 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1501 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1561 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1621 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1681 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1741 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1801 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1861 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1921 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1981 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2041 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2101 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     2161 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2221 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2281 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2341 atggaaaaac gccttcttct tgaactcgaa ttgaggcgcg cccgaagtta tctagtagca
     2401 ggtcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag
     2461 cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg
     2521 agactctagc gagcgccgga gtataaatag aggcgcttcg tctacggagc gacaattcaa
     2581 ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc
     2641 tgaacaagct aaacaatctg cagtaaagtg caagttaaag tgaatcaatt aaaagtaacc
     2701 agcaaccaag taaatcaact gcaactactg aaatctgcca agaagtaatt attgaataca
     2761 agaagagaac tctgaatagg gaattgggaa ttcgagatct aaagagcctg ctaaagcaaa
     2821 aaagaagtca ccatgtcgac cggctcgagc tacccctacg acgtgcccga ctacgcagct
     2881 ggtagctacc cctacgacgt gcccgactac gcatcgctag ctagatatcg cggtacctgt
     2941 ctagacctgc aggatgcggc cgcacactag taggcctgta gagtttccca tttaataatt
     3001 catattatct cgaatctagt caattacggc tttcctcaaa tagaaaaata aaaaaaatga
     3061 aaaaatgcac ttgccattta aacttagacg cgataacgtt caattcgagt tcaagaagaa
     3121 ggcgtttttc cataggctcc gcccctagat aacttcgtat aatgtatgct atacgaagtt
     3181 atcatgccgg ggatctaatt caattagaga ctaattcaat tagagctaat tcaattagga
     3241 tccaagctta tcgatttcga accctcgacc gccggagtat aaatagaggc gcttcgtcta
     3301 cggagcgaca attcaattca aacaagcaaa gtgaacacgt cgctaagcga aagctaagca
     3361 aataaacaag cgcagctgaa caagctaaac aatcgggcgg ccgtactaga gccggtcgcc
     3421 accatggtgt cgagcgcaag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3481 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3541 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3601 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3661 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3721 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3781 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3841 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3901 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3961 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4021 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     4081 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4141 ctgtacaagt actcagatct tgcaagcttg tagagtttcc catttaataa ttcatattat
     4201 ctcgaatcta gtcaattacg gctttcctca aatagaaaaa taaaaaaaat gaaaaaatgc
     4261 acttgccatt taaacttaga cgcgataagc ttgatataac ttcgtataat gtatgctata
     4321 cgaagttata cgcgtactag a
//
