LOCUS       sequence-251761-        5547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of mCherry-CaaX in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5547)
  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 5547)
  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 mCherry-CaaX
FEATURES             Location/Qualifiers
     source          1..5547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     143..165
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     157..174
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     158..174
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        180..606
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             638..1345
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(783..801)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1051..1070
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1436..1453)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1455..1589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             2064..2663
                     /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(2064..2083)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2560..2580
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2796..2812)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2797..2813)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2828..2962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3105..3121)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3105..3121)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3118..3140)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3129..3145
                     /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(3153..3159,3160..3177,3178..3183))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3198..3219
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3336..3353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3507..4095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3587..3606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4266..5057,5058..5126))
                     /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     4889..4908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5127..5231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5299..5317
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5355..5377)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5477..5496
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
       61 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
      121 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatta
      181 attcgttgca ggacaggatg tggtgcccga tgtgactagc tctttgctgc aggccgtcct
      241 atcctctggt tccgataaga gacccagaac tccggccccc caccgcccac cgccaccccc
      301 atacatatgt ggtacgcaag taagagtgcc tgcgcatgcc ccatgtgccc caccaagagc
      361 tttgcatccc atacaagtcc ccaaagtgga gaaccgaacc aattcttcgc gggcagaaca
      421 aaagcttctg cacacgtctc cactcgaatt tggagccggc cggcgtgtgc aaaagaggtg
      481 aatcgaacga aagacccgtg tgtaaagccg cgtttccaaa atgtataaaa ccgagagcat
      541 ctggccaatg tgcatcagtt gtggtcagca gcaaaatcaa gtgaatcatc tcagtgcaac
      601 taaagggggg atctagatcg gggtacctac tagtcaaatg gtgagcaagg gcgaggagga
      661 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
      721 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      781 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      841 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
      901 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      961 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     1021 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     1081 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     1141 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     1201 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     1261 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     1321 cggcggcatg gacgagctgt acaagctcga gaaaatgtcc aaggatggta agaaaaagaa
     1381 gaagaagtca aaaaccaagt gtgttatcat gtaagtttaa acgcccgctg atcagcctcg
     1441 actgtgcctt ctaagatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     1501 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1561 taaccattat aagctgcaat aaacaagtta actctagagg atccggtgcg tggtggtttc
     1621 atgcttctgg gaacggcaaa tgggtttagg attgggaacc cctcatcatc tgttggaata
     1681 tactattcaa cctacaaaag taacgttaaa caacactact ttatatttga tatgaatggc
     1741 cacacctttt atgccataaa acatattgta agagaatacc actcttttta ttccttcttt
     1801 ccttcttgta cgttttttgc tgtaagtagg tcgtggtgct ggtgttgcag ttgaaataac
     1861 ttaaaatata aatcataaaa ctcaaacata aacttgacta tttatttatt tattaagaaa
     1921 ggaaatataa attataaatt acaacagatc ccccgggctg caggaattcg atagcttgca
     1981 tgcctgcagg tcggccgcca cgaccggtgc cgccaccatc ccctgaccca cgcccctgac
     2041 ccctcacaag gagacgacct tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     2101 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     2161 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     2221 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     2281 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     2341 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     2401 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     2461 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     2521 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     2581 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     2641 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
     2701 gcgcacgacc ccatggctcc gaccgaagcc acccggggcg gccccgccga ccccgcaccc
     2761 gcccccgagg cccaccgact ctagatcaag cttatcgata ccgtcgacct cgaggccagc
     2821 tagcttgaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2881 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2941 tgtatcttat catgtctgga tcgggtaccc aattgcattc attttatgtt tcaggttcag
     3001 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     3061 tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
     3121 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3181 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3241 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3301 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3361 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3421 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3481 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3541 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3601 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3661 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3721 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3781 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3841 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3901 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3961 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4021 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4081 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4141 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4201 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4261 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4321 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4381 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4441 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4501 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4561 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4621 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4681 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4741 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4801 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4861 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4921 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4981 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5041 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5101 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5161 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5221 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5281 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     5341 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5401 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5461 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5521 gaaataccgc acagatgcgt aaggaga
//
