LOCUS       sequence-135057-        3666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gap-repair recombineering for Drosophila transgenesis, contains loxP
            cassette with GFP, Amp resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3666)
  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 3666)
  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
FEATURES             Location/Qualifiers
     source          1..3666
                     /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    2474..2507
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2743..2752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2764..3474
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             2764..3474
                     /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(2803..2824)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3064..3083)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3411..3432
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    3621..3654
                     /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 ttctgtcgac agctgcagta ctagtcaagc
     2401 ttaggatccg atatccatgg tgctagcgag atcttcctcg agggtaccgg cggccgcgtg
     2461 cggccgcgtc tagataactt cgtataatgt atgctatacg aagttatcat gccggggatc
     2521 taattcaatt agagactaat tcaattagag ctaattcaat taggatccaa gcttatcgat
     2581 ttcgaaccct cgaccgccgg agtataaata gaggcgcttc gtctacggag cgacaattca
     2641 attcaaacaa gcaaagtgaa cacgtcgcta agcgaaagct aagcaaataa acaagcgcag
     2701 ttgaacaagc taaacaatcg ggcggccgta ctagagccgg tcgccaccat ggtgtcgagc
     2761 gcaagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2821 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2881 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2941 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     3001 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3061 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3121 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3181 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     3241 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3301 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3361 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3421 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtactca
     3481 gatcttgcaa gcttgtagag tttcccattt aataattcat attatctcga atctagtcaa
     3541 ttacggcttt cctcaaatag aaaaataaaa aaaatgaaaa aatgcacttg ccatttaaac
     3601 ttagacgcga taagcttgat ataacttcgt ataatgtatg ctatacgaag ttatacgcgt
     3661 actaga
//
