LOCUS       sequence-258858-        5786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains ~1kb homology regions flanking Drosophila crk locus. Used 
            to make crk null mutant. Homology regions cloned into AarI and SapI 
            sites of pHD-dsRed-attP (Harrison, O'Connor-Giles, Wildonger)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5786)
  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 5786)
  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: pHD-dsRed-attP_dCrk_flankingHRs
FEATURES             Location/Qualifiers
     source          1..5786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     159..178
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1238..1271
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1497..1506
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1503..2180
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(1633..1651)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1683..1703)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1901..1920
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2095..2118
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2301..2422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2338..2357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2392..2411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    2429..2462
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3930..3947)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3930..3946)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4045..4062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4216..4804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4296..4315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4926..5717,5718..5786))
                     /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     5549..5568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc
      121 aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac accccttgtt
      181 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
      241 gccgtagcgc cgatggtagt gtggggactc cccatgcgag agtagggaac tgccaggcat
      301 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gcccgggcta attatggggt
      361 gtcgcccttc gccttgagca tgcaaaggaa tggctattac aatataattt gattagtatg
      421 ctacgcaaac ctctagcatt ttgataggaa aaagattcgg acaaagattc gttgtcagtt
      481 ttttgaagaa gatgaagagg cggatggtgc agtggtctgg ccatgtgagg aaagtttatt
      541 tcccacgggc ccctttttcc tatttttgat cctagcttca aaatcttata tatcttatgt
      601 ccatagcgta agataagtta aatttttcca cttttatatt acttttatat tatcgacttt
      661 tgttttgtct tgcatataag accatacatc agatgatttt tgatcaggtg aaaaccgaga
      721 aacgaaagtt tgtttcttaa gtggaatcac cgcgagtggt tttggccctg cagattgtat
      781 ttctgaagtg ccaacttgtg ccgggagtcc ggactcaatg ttcccttgtg gtggtaaact
      841 aatcgggaca ggtagaatca ctggttaaga aaccagtgca gacaattcgg aatctttagc
      901 aatcccaggg tgggctccct ccacagcgga ttgatcggat tgctccaaat cttatttagc
      961 tgccgatctt agcaacatat gcggattttc tacgattgac agctgatcag ttgtggagtc
     1021 aaaataacat gtcaaccaga tatattcgta ttcctaatat tcctgtaaat atccttgaat
     1081 tgaacagaag aacaaatata tcggttttac ttaaaaatta gaatatcaaa ttgatatgca
     1141 ttgatcacgg ttttacattt agattttaaa agctgacagt acgtcctgta gtgccccaac
     1201 tggggtaacc tttgagttct ctcagttggg ggcgtagata acttcgtata atgtatgcta
     1261 tacgaagtta tcgtacggga tctaattcaa ttagagacta attcaattag agctaattca
     1321 attaggatcc aagcttatcg atttcgaacc ctcgaccgcc ggagtataaa tagaggcgct
     1381 tcgtctacgg agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag
     1441 ctaagcaaat aaacaagcgc agctgaacaa gctaaacaat cggctcgaag ccggtcgcca
     1501 ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg
     1561 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1621 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     1681 tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc gccgacatcc
     1741 ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1801 aggacggcgg cgtggtgacc gtgacccagg actcctccct ccaggacggc tccttcatct
     1861 acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg cagaagaaga
     1921 ctatgggctg ggaggcgtcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     1981 agatccacaa ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagtcca
     2041 tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac tccaagctgg
     2101 acatcacctc ccacaacgag gactacacca tcgtggagca gtacgagcgc gccgagggcc
     2161 gccaccacct gttcctgtag gggccgcgac tctagatcat aatcagccat accacatttg
     2221 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2281 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     2341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2401 ccaaactcat caatgtatct taaccggtat aacttcgtat aatgtatgct atacgaagtt
     2461 attgctacag aaaacccttt gggccttata ttttttatac ccgttactcg tagagtaaaa
     2521 gggaatacta caataggttc gttgaaaagt atgtaacagg tggaagaaag cgcttctgac
     2581 catataaagg ctgttcaata tacaaagacc ccaaaagccg gatacatctc cgagtaacgc
     2641 cagaacgcca gaagtttttt tatcacccgc aaagaggtcc ttatttggcc cttcgaccac
     2701 cttccaaaat ttgatatttc caaacgtcca taattcgagg ctaacagaac ctacctctaa
     2761 ggcaccaact ctacaagctg cccaaactaa tttgacgtcg aatccttaat caccgacgca
     2821 tcaccaacgc agtattaaaa ctatgacata tactttgcag catcatatga tttcattcat
     2881 gtcgttcgtg tggattatca cgcaagacct atcttcctgg aacgttaaaa taatgtgtat
     2941 ctttagatca atagttagtt agtgcttctc ctgataccac gttccgagag cacctagagt
     3001 acgcccacaa aaaggccagc gctactactg gttctctgcc gagaatgctt ctaaacacga
     3061 gaggtcctaa gcaaaccact agctaccacg ttgataacgg gcgacaaagc atttttgggc
     3121 ggctgaaggg cgttagagtg gccgtggcaa aaagtttttt gggaaatcga tagaaaatta
     3181 caagactaat caaaatattt aaagaatatc aacacacttt ttcaaaagtg cgggcgtggc
     3241 agttttgcgc ggtttgtagg cgttagagtg ggcgtggcaa catgggtcaa cgctgcgctg
     3301 cgtcttttct agattctgta tgctgaatct catccttcta gcttttatag ttcctgtaat
     3361 cgtgacgttc atacggacag acggacatga ccagatcgtc tcggctattg atcctgatca
     3421 ggaatggcaa catgggtcaa caaacttgtg ctgtctatgt tcgtcaatcg agttcaaggg
     3481 cgacacaaaa tttattctaa atgcataata aatactgata acatcttata gtttgtatta
     3541 tattttgtat tatcgttgac atgtataatt ttgatatcaa aaactgattt tccctttatt
     3601 attttcgaga tttattttct taattctctt taacaaacta gaaatattgt atatacaaaa
     3661 aatcataaat aatagatgaa tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt
     3721 atcttattta aagtgcgttg cttttttctc atttataagg ttaaataatt ctcatatatc
     3781 aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc tctttcccta aactcccccc
     3841 ataaaaaaac ccgccgaagc gggtttttac gttatttgcg gattaacgat tactcgttat
     3901 cagaaccgcc cagggggccc gagcttaaga ctggccgtcg ttttacaaca cagaaagagt
     3961 ttgtagaaac gcaaaaaggc catccgtcag gggccttctg cttagtttga tgcctggcag
     4021 ttccctactc tcgccttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4081 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4141 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4201 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4261 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4321 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4381 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4441 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4501 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4561 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4621 cttgaagtgg tgggctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4681 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4741 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4801 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgcgcgt
     4861 aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc
     4921 tctgcttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4981 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5041 ggccccagcg ctgcgatgat accgcgagaa ccacgctcac cggctccgga tttatcagca
     5101 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5161 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5221 cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5281 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5341 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5401 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5461 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5521 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5581 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5641 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5701 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5761 gcgacacgga aatgttgaat actcat
//
