LOCUS       sequence-251716-        6330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Crk-mCherry in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  AUTHORS   Williamson AP, Vale RD
  TITLE     Spatial control of Draper receptor signaling initiates apoptotic 
            cell engulfment.
  JOURNAL   J Cell Biol. 2018 Nov 5;217(11):3977-3992. doi: 
            10.1083/jcb.201711175. Epub 2018 Aug 23.
  PUBMED    30139739
REFERENCE   2  (bases 1 to 6330)
  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"; doi: 
            "10.1083/jcb.201711175"; journalName: "J Cell Biol"; date: 
            "2018-11-5- 5"; volume: "217"; issue: "11"; pages: "3977-3992"
COMMENT     Sequence Label: pMT Crk-mCherry
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     129..151
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     143..160
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     144..160
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        166..592
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             1473..2180
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1615..1633)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1883..1902
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2205..2222)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2224..2358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2833..3432
                     /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(2833..2852)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3329..3349
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3565..3581)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3566..3582)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3597..3731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3874..3890)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3874..3890)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3887..3909)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3898..3914
                     /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)."
     promoter        complement(join(3922..3928,3929..3946,3947..3952))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3967..3988
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4105..4122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4276..4864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4356..4375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5035..5826,5827..5895))
                     /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     5658..5677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5896..6000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6068..6086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6124..6146)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6246..6265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
       61 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      121 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattaattc gttgcaggac
      181 aggatgtggt gcccgatgtg actagctctt tgctgcaggc cgtcctatcc tctggttccg
      241 ataagagacc cagaactccg gccccccacc gcccaccgcc acccccatac atatgtggta
      301 cgcaagtaag agtgcctgcg catgccccat gtgccccacc aagagctttg catcccatac
      361 aagtccccaa agtggagaac cgaaccaatt cttcgcgggc agaacaaaag cttctgcaca
      421 cgtctccact cgaatttgga gccggccggc gtgtgcaaaa gaggtgaatc gaacgaaaga
      481 cccgtgtgta aagccgcgtt tccaaaatgt ataaaaccga gagcatctgg ccaatgtgca
      541 tcagttgtgg tcagcagcaa aatcaagtga atcatctcag tgcaactaaa ggggggatct
      601 agatcggggt acctactagt caaatggata catttgacgt ttctgatagg aacagctggt
      661 actttggtcc catgtctaga caggatgcta ctgaagtttt gatgaacgaa cgcgagcggg
      721 gagtgttttt agtccgtgat agtaactcga tagcagggga ttatgtactt tgtgtaagag
      781 aagatacaaa agttagcaac tacatcatta acaaagttca acaacaggat caaatcgttt
      841 accgcattgg ggatcagtct tttgacaatc taccgaaact cttaactttt tacactcttc
      901 attatttgga tacaacccct ttaaaacggc ctgcgtgtag aagggtggaa aaagtaatag
      961 gaaagttcga tttcgttggc agcgatcaag atgatttacc ttttcaaaga ggtgaagttt
     1021 taacaatagt tcgaaaagac gaggatcaat ggtggactgc gcgtaactcc tcggggaaaa
     1081 ttggtcaaat accggttccc tatatacaac agtatgacga ttatatggat gaagatgcta
     1141 ttgataaaaa cgaaccttcc atttcgggat ctagcaatgt atttgaaagt actcttaaaa
     1201 ggacagattt aaatcgaaaa ctacctgcat acgcccgcgt aaaacagtca agggtcccta
     1261 acgcatacga taagactgca ttaaaattgg aaataggtga cattattaaa gtcactaaaa
     1321 caaacattaa tgggcaatgg gagggagaat taaatggaaa aaatggtcat tttcccttca
     1381 cgcacgttga atttgtcgat gattgtgatt taagcaaaaa ctccacagaa atatgccggc
     1441 gcgccggcgg ccgctcgagt ctagagggcc ccgtgagcaa gggcgaggag gataacatgg
     1501 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     1561 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     1621 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     1681 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     1741 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     1801 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     1861 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     1921 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     1981 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     2041 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     2101 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     2161 tggacgagct gtacaagtaa taggtttaaa cgcccgctga tcagcctcga ctgtgccttc
     2221 taagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2281 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2341 agctgcaata aacaagttaa ctctagagga tccggtgcgt ggtggtttca tgcttctggg
     2401 aacggcaaat gggtttagga ttgggaaccc ctcatcatct gttggaatat actattcaac
     2461 ctacaaaagt aacgttaaac aacactactt tatatttgat atgaatggcc acacctttta
     2521 tgccataaaa catattgtaa gagaatacca ctctttttat tccttctttc cttcttgtac
     2581 gttttttgct gtaagtaggt cgtggtgctg gtgttgcagt tgaaataact taaaatataa
     2641 atcataaaac tcaaacataa acttgactat ttatttattt attaagaaag gaaatataaa
     2701 ttataaatta caacagatcc cccgggctgc aggaattcga tagcttgcat gcctgcaggt
     2761 cggccgccac gaccggtgcc gccaccatcc cctgacccac gcccctgacc cctcacaagg
     2821 agacgacctt ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     2881 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     2941 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3001 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3061 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3121 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     3181 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3241 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     3301 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3361 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     3421 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     3481 catggctccg accgaagcca cccggggcgg ccccgccgac cccgcacccg cccccgaggc
     3541 ccaccgactc tagatcaagc ttatcgatac cgtcgacctc gaggccagct agcttgaact
     3601 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3661 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3721 atgtctggat cgggtaccca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3781 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     3841 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3901 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3961 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4021 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4081 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4141 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4201 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4261 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4321 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4381 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4441 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4501 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4561 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4621 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4681 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4741 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4801 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4861 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4921 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4981 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5041 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5101 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5161 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5221 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5281 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5341 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5401 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5461 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5521 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5581 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5641 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5701 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5761 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5821 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5881 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5941 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6001 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     6061 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6121 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     6181 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     6241 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     6301 cagatgcgta aggagaaaat accgcatcag
//
