LOCUS       sequence-258814-        8261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rescue construct for targeting into crk[Delta-attP]. Contains 110bp 
            upstream of transcription start site, mNeonGreen, 3XFLAG, and dCrk 
            cDNA, 3' UTR and 99bp downstream..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8261)
  AUTHORS   Spracklen AJ, Thornton-Kolbe EM, Bonner AN, Florea A, Compton PJ, 
            Fernandez-Gonzalez R, Peifer M
  TITLE     The Crk adapter protein is essential for Drosophila embryogenesis, 
            where it regulates multiple actin-dependent morphogenic events.
  JOURNAL   Mol Biol Cell. 2019 Jul 18:mbcE19050302. doi: 
            10.1091/mbc.E19-05-0302.
  PUBMED    31318326
REFERENCE   2  (bases 1 to 8261)
  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: "Mol Biol 
            Cell. 2019 Jul 18:mbcE19050302. doi: 10.1091/mbc.E19-05-0302."
COMMENT     Sequence Label: pGE-attB-GMR_mNeonGreen::3XFLAG::dCrk
FEATURES             Location/Qualifiers
     source          1..8261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..31,32..233)
                     /label=P element 3' end
                     /note="P element 3' end"
     protein_bind    353..386
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1453..1518
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    2589..2622
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(5634..6188,6189..6219)
                     /label=P element 5' end
     rep_origin      complement(6454..7042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6534..6553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7213..8004,8005..8073))
                     /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     7836..7855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8074..8178)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgatgaaa taacataagg tggtcccgtc ggcaagagac atccacttaa cgtatgcttg
       61 caataagtgc gagtgaaagg aatagtattc tgagtgtcgt attgagtctg agtgagacag
      121 cgatatgatt gttgattaac ccttagcatg tccgtggggt ttgaattaac tcataatatt
      181 aattagacga aattattttt aaagttttat ttttaataat ttgcgagtac gcaaagctag
      241 agttgttggt tggcacacca caaatatact gttgccgagc acaattccgc tcgaattgcc
      301 ggctgtacac ttaagaagtt cctactgcag gtaatgaatt gccgcggtgc gggtgccagg
      361 gcgtgccctt gggctccccg ggcgcgtact ccacgaattc aaaggggttt accaccacag
      421 agcagttgct cgaaggagtt cttttcaatc gaatgtgccc tgtgcaagtg ccgcattacg
      481 gttctacggt acaatattgg cataaaactt ccaagtggaa ataatcgatc aaacattatc
      541 gatagtgttt ctatgtgtgg cccagccaga tacacatata aaaggcaaat gttttaaggc
      601 cagaaattcg aaaatgaaat cactgaagaa ttctggtata taataggctg tatgctatat
      661 ttgcattata tcacataatt tctatttatt taatctggac ggaatttgtt gtaatttgtc
      721 taagataaac cttaaacgga aatggtttcc aaaggtgaag aagacaatat ggcttccctg
      781 cctgcaaccc acgagttgca catttttgga tcgatcaacg gcgtcgattt tgatatggta
      841 ggtcagggaa cgggaaaccc taatgacggt tatgaagaac tcaatctgaa gtcgacaaag
      901 ggtgatctcc aattttcgcc ttggattctc gtaccgcaca ttggatatgg atttcatcaa
      961 tatttgccat atccagacgg aatgtccccg ttccaggcgg ctatggtcga tggatccggt
     1021 taccaggtcc acaggacaat gcagttcgag gacggcgcat cgctcacggt taattaccgt
     1081 tacacgtacg agggtagtca tataaagggc gaggcacagg tgaaaggaac cggattcccc
     1141 gccgatggcc cggttatgac caattcgctc actgcggctg attggtgtcg gtccaaaaaa
     1201 acatatccta acgacaaaac tataatatcc accttcaaat ggtcctacac gaccggaaat
     1261 ggaaagcgtt atcgttcgac ggctcggacc acttatactt tcgctaagcc catggcggca
     1321 aactacctca agaatcagcc tatgtatgta ttccgaaaaa cagagctcaa gcacagtaaa
     1381 acggagctga atttcaagga gtggcaaaaa gctttcacgg acgttatggg catggatgaa
     1441 ctctacaaga tggactacaa ggatcatgac ggtgactaca aagaccatga tatagattac
     1501 aaggatgacg acgacaagat ggatacattt gacgtttctg ataggaacag ctggtacttt
     1561 ggtcccatgt ctagacagga tgctactgaa gttttgatga acgaacgcga gcggggagtg
     1621 tttttagtcc gtgatagtaa ctcgatagca ggggattatg tactttgtgt aagagaagat
     1681 acaaaagtta gcaactacat cattaacaaa gttcaacaac aggatcaaat cgtttaccgc
     1741 attggggatc agtcttttga caatctaccg aaactcttaa ctttttacac tcttcattat
     1801 ttggatacaa cccctttaaa acggcctgcg tgtagaaggg tggaaaaagt aataggaaag
     1861 ttcgatttcg ttggcagcga tcaagatgat ttaccttttc aaagaggtga agttttaaca
     1921 atagttcgaa aagacgagga tcaatggtgg actgcgcgta actcctcggg gaaaattggt
     1981 caaataccgg ttccctatat acaacagtat gacgattata tggatgaaga tgctattgat
     2041 aaaaacgaac cttccatttc gggatctagc aatgtatttg aaagtactct taaaaggaca
     2101 gatttaaatc gaaaactacc tgcatacgcc cgcgtaaaac agtcaagggt ccctaacgca
     2161 tacgataaga ctgcattaaa attggaaata ggtgacatta ttaaagtcac taaaacaaac
     2221 attaatgggc aatgggaggg agaattaaat ggaaaaaatg gtcattttcc cttcacgcac
     2281 gttgaatttg tcgatgattg tgatttaagc aaaaactcca cagaaatatg ctaaatagga
     2341 aggaatggaa ggaatttctt ttgataattt tgatttttga gctaattgta tgattaataa
     2401 gttgtgcata ccaattgtta acataatcag aaaattaaaa tcttattaag taaaataaaa
     2461 ataaacgggt acgaaggaca taataaaatt tttaccgttt tttcgtatga ttcttataca
     2521 aaaaaattat gaatttgaaa atcgatttgg ttttttttat gaattgtgta actaaacggt
     2581 agggtaccat aacttcgtat aatgtatgct atacgaagtt atcgtacgac tagcacctga
     2641 ctgtcgagcc cagtggaaac ccttgaaatg cctttaagtc gagcccagtg gaaacccttg
     2701 aaatgccttt aagtcgagcc cagtggaaac ccttgaaatg cctttaagtc gagcccagtg
     2761 gaaacccttg aaatgccttt aagtcgagcc cagtggaaac ccttgaaatg cctttaagtc
     2821 gagtagcagt cgaggtaggt acgccatgcc ccgttattct ctattcgttt tgtgactctc
     2881 cctctctgta ctattgctct ctcactctgt cgcacagtaa acggcacgct attctcgttg
     2941 cttcgagaga gcgcgcctcg aatgttcgcg aaaagagcgc cggagtataa atagaggcgc
     3001 ttcgtcgacg gagcgtcaat tcaattcaaa caagcaaagt gaacacatcg cgaagcgtaa
     3061 gctgagcaaa caaacaagcg cagctgaaca agctaaacaa tctgcaataa agtgcaagtt
     3121 aaagtgaatc aattaaaagt aaccaacaac caagtaatta aactaaaaac tgcaactact
     3181 gaaatcaacc aagaagtcat tattgaagac aagaagagaa ctctgaatag gtcgatagcg
     3241 tcaattcggc acgagcggtt tcgtgacgaa gctccaagcg gtttacgcca tcaattaaac
     3301 acaaagtgct gtgccaaaac tcctctcgct tcttattttt gtttgttttt tgagtgattg
     3361 ggtgggtgat tggttttggg tgggtaagca ggggaaagtg tgaaaaatcc cggcaatggg
     3421 ccaagaggat caggagctat taattcgcgg aggcagcaaa cacccatctg ccgagcatct
     3481 gaacaatggt gacagcggag cggcttcgca gagctgcatt aaccagggct tcgggcaggc
     3541 caaaaactac ggcacgctcc tgccacccag tccgccggag gactccggtt cagggagcgg
     3601 ccaactagcc gagaacctca cctatgcctg gcacaatatg gacatctttg gggcggtcaa
     3661 tcagccgggc tccggatggc ggcagctggt caaccggaca cgcggactat tctgcaacga
     3721 gcgacacata ccggcgccca ggaaacattt gctcaagaac gtttgcggcg tggcctatcc
     3781 gggcgaactt ttggccgtga tgggcagttc cggtgccgga aagacgaccc tgctgaatgc
     3841 ccttgccttt cgatcgccgc agggcatcca agtatcgcca tccgggatgc gactgctcaa
     3901 tggccaacct gtggacgcca aggagatgca ggccaggtgc gcctatgtcc agcaggatga
     3961 cctctttatc ggctccctaa cggccaggga acacctgatt ttccaggcca tggtgcggat
     4021 gccacgacat ctgacctatc ggcagcgagt ggcccgcgtg gatcaggtga tccaggagct
     4081 ttcgctcagc aaatgtcagc acacgatcat cggtgtgccc ggcagggtga aaggtctgtc
     4141 cggcggagaa aggaagcgtc tggcattcgc ctccgaggca ctaaccgatc cgccgcttct
     4201 gatctgcgat gagcccacct ccggactgga ctcatttacc gcccacagcg tcgtccaggt
     4261 gctgaagaag ctgtcgcaga agggcaagac cgtcatcctg accattcatc agccgtcttc
     4321 cgagctgttt gagctctttg acaagatcct tctgatggcc gagggcaggg tagctttctt
     4381 gggcactccc agcgaagccg tcgacttctt ttcctacgtg ggtgcccagt gtcctaccaa
     4441 ctacaatccg gcggactttt acgtacaggt gttggccgtt gtgcccggac gggagatcga
     4501 gtcccgtgat cggatcgcca agatatgcga caattttgcc attagcaaag tagcccggga
     4561 tatggagcag ttgttggcca ccaaaaattt ggagaagcca ctggagcagc cggagaatgg
     4621 gtacacctac aaggccacct ggttcatgca gttccgggcg gtcctgtggc gatcctggct
     4681 gtcggtgctc aaggaaccac tcctcgtaaa agtgcgactt attcagacaa cgatggttgc
     4741 catcttgatt ggcctcatct ttttgggcca acaactcacg caagtgggcg tgatgaatat
     4801 caacggagcc atcttcctct tcctgaccaa catgaccttt caaaacgtct ttgccacgat
     4861 aaatgtgttc acctcagagc tgccagtttt tatgagggag gcccgaagtc gactttatcg
     4921 ctgtgacaca tactttctgg gcaaaacgat tgccgaattg ccgctttttc tcacagtgcc
     4981 actggtcttc acggcgattg cctatccgat gatcggactg cgggccggag tgctgcactt
     5041 cttcaactgc ctggcgctgg tcactctggt ggccaatgtg tcaacgtcct tcggatatct
     5101 aatatcctgc gccagctcct cgacctcgat ggcgctgtct gtgggtccgc cggttatcat
     5161 accattcctg ctctttggcg gcttcttctt gaactcgggc tcggtgccag tatacctcaa
     5221 atggttgtcg tacctctcat ggttccgtta cgccaacgag ggtctgctga ttaaccaatg
     5281 ggcggacgtg gagccgggcg aaattagctg cacatcgtcg aacaccacgt gccccagttc
     5341 gggcaaggtc atcctggaga cgcttaactt ctccgccgcc gatctgccgc tggactacgt
     5401 gggtctggcc attctcatcg tgagcttccg ggtgctcgca tatctggctc taagacttcg
     5461 ggcccgacgc aaggagtagc cgacatatat ccgaaataac tgtttgtttt ttttttttac
     5521 cattattacc atcgtgttta ctgtttattg ccccctcaaa aagctaatgt aattatattt
     5581 gtgccaataa aaacaagata tgacctatag aatacaaaga tccaaaccta gtggctgctg
     5641 ctctaaacga cgcatttcgt actccaaagt acgaattttt tccctcaagc tcttattttc
     5701 attaaacaat gaacaggacc taacgcacag tcacgttatt gtttacataa atgatttttt
     5761 ttactattca aacttactct gtttgtgtac tcccactggt atagccttct tttatctttt
     5821 ctggttcagg ctctatcact ttactaggta cggcatctgc gttgagtcgc ctccttttaa
     5881 atgtctgacc ttttgcaggt gcagccttcc actgcgaatc attaaagtgg gtatcacaaa
     5941 tttgggagtt ttcaccaagg ctgcacccaa ggctctgctc ccacaatttt ctcttaatag
     6001 cacacttcgg cacgtgaatt aattttactc cagtcacagc tttgcagcaa aatttgcaat
     6061 atttcatttt tttttattcc acgtaagggt taatgttttc aaaaaaaaat tcgtccgcac
     6121 acaacctttc ctctcaacaa gcaaacgtgc actgaattta agtgtatact tcggtaagct
     6181 tcggctatcg acgggaccac cttatgttat ttcatcatgg gccagaccca cgtagtccag
     6241 cggcagatcg gcggcggaga agttaagcgt ctccaggatg accttgcccg aactggggca
     6301 cgtggtgttc gacgatgtgc agctaatttc gcccggctcc acgtccgccc attggttaat
     6361 cagcagaccc tcgttggcgt aacggaacca tgagaggtac gacaaccatt tgaggtatac
     6421 tggcaccgag cccgagttca agaagaaggc gtttttccat aggctccgcc cccctgacga
     6481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctcgaagtc ttagagccag
     8221 atatgcgagc acccggaagc tcacgatgag aatggccaga c
//
