LOCUS       sequence-251766-        8680 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Inducible expression of Draper-mCherry in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8680)
  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 8680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 Draper-mCherry
FEATURES             Location/Qualifiers
     source          1..8680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..24
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     218..240
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     232..249
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     233..249
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        255..681
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             716..733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3917..4624
                     /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(4059..4077)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4327..4346
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4644..4661)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4663..4797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5272..5871
                     /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(5272..5291)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5768..5788
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6004..6020)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6005..6021)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6036..6170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6313..6329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6313..6329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6326..6348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6337..6353
                     /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(6361..6367,6368..6385,6386..6391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6406..6427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6544..6561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6715..7303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6795..6814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7474..8265,8266..8334))
                     /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     8097..8116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8335..8439)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8507..8525
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8563..8585)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
       61 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
      121 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      181 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      241 acggccagtg aattaattcg ttgcaggaca ggatgtggtg cccgatgtga ctagctcttt
      301 gctgcaggcc gtcctatcct ctggttccga taagagaccc agaactccgg ccccccaccg
      361 cccaccgcca cccccataca tatgtggtac gcaagtaaga gtgcctgcgc atgccccatg
      421 tgccccacca agagctttgc atcccataca agtccccaaa gtggagaacc gaaccaattc
      481 ttcgcgggca gaacaaaagc ttctgcacac gtctccactc gaatttggag ccggccggcg
      541 tgtgcaaaag aggtgaatcg aacgaaagac ccgtgtgtaa agccgcgttt ccaaaatgta
      601 taaaaccgag agcatctggc caatgtgcat cagttgtggt cagcagcaaa atcaagtgaa
      661 tcatctcagt gcaactaaag gggggatcta gatcggggta cctactagtg aaatgcacca
      721 tcaccatcat cacgaagatc aagttgaccc aagacttatt gacggaaaag gaggaggagg
      781 ccggcccatg ttgccggtaa tcctcatagc gtgcctggcc cagctggtac tggctcaggc
      841 ggatcttaaa gatctggatg gacccaatat ctgcaaaaga agagaactat ataacgtaga
      901 cgttgtctac acagaactgc aatcgttcca ggaacggggc tccacctggt gcgtcacatt
      961 cccgccgagg tgctccacct atcgcattaa acaccgagtg gtcaacaaaa ccaagacgat
     1021 tgcgaagaac cgtattgtga gggactgttg tgatggctac atcgcgagcg caggagaatg
     1081 cgtgccgcat tgttcggaac cctgccagca cggtagatgc atctccccgg agaagtgcaa
     1141 gtgtgatcat ggttacggag gacccgcctg cgatataaat tgcccacccg gctggtatgg
     1201 ccggaactgc tcgatgcagt gcgactgcct gaacaacgcg gtatgtgagc ccttttcagg
     1261 cgactgcgag tgcgccaagg gctacactgg agcacgctgc gcggacatct gtccggaggg
     1321 ctttttcggg gccaactgct cggagaagtg ccgatgcgag aacggcggca agtgtcacca
     1381 cgtgtccggc gaatgccagt gtgctcccgg tttcacgggt cccctatgtg atatgcgctg
     1441 tccggatgga aagcatggcg cccagtgcca gcaggactgt ccctgccaga acgacggaaa
     1501 gtgtcagcca gagaccgggg cctgtatgtg caatccaggc tggacgggcg acgtgtgtgc
     1561 aaacaagtgt ccggtgggca gctacggccc tggatgccaa gagagctgcg agtgctacaa
     1621 gggagcgccc tgccaccaca tcaccggcca gtgcgagtgt ccgcccggat atcgcggaga
     1681 acgctgcttc gacgagtgtc agctgaacac gtacggattc aactgcagca tgacctgcga
     1741 ctgcgcgaac gatgccatgt gcgatcgggc gaacggaacg tgcatctgca accccggctg
     1801 gacgggtgcc aagtgcgccg agcggatctg cgaggcgaac aagtacgggc tggactgtaa
     1861 ccgcacctgc gagtgcgaca tggagcacac ggatctgtgt cacccggaga cgggaaattg
     1921 ccagtgcagc atcggatgga gcagtgctca gtgcaccaga ccctgcacct tcctgcgcta
     1981 cggcccgaat tgcgagctga cctgcaactg caagaacgga gccaagtgct cgccggtcaa
     2041 tggtacgtgc ctgtgtgcgc cgggctggcg cggacccacc tgcgaggaga gctgcgagcc
     2101 gggcaccttt ggtcaggact gcgccctgcg ctgcgactgc cagaacggag ccaagtgcga
     2161 gccggagacg ggccagtgct tgtgcacggc cggatggaag aacatcaagt gcgaccggcc
     2221 gtgtgatctg aaccacttcg gacaggactg cgccaaggtg tgcgactgcc acaataacgc
     2281 cgcctgcaat ccgcagaacg gaagctgcac ctgtgccgcc ggctggacgg gcgagcggtg
     2341 cgaaaggaag tgcgataccg gcaaattcgg acacgactgc gcccagaagt gccagtgcga
     2401 cttcaacaac agcctggcct gcgatgccac caatggaaga tgcgtgtgca agcaggattg
     2461 gggaggcgtt cattgcgaga ccaactgtcg gagcggttac tatggtgaaa attgtgataa
     2521 agtttgcaga tgcctgaata actcctcctg cgaccccgat tccggtaact gcatctgctc
     2581 agccggttgg actggcgccg actgtgcgga accctgccca cctggattct acggcatgga
     2641 atgcaaggaa cggtgcccgg agatcctcca tggcaacaag agctgcgatc acatcacagg
     2701 tgaaattctc tgccgcacgg gttacatagg tctcacctgc gagcacccat gtcctgctgg
     2761 actctacgga cccggctgca agctcaagtg caattgcgag cacggcggcg agtgcaacca
     2821 cgttacaggc cagtgccagt gcctgccagg gtggacaggt tccaactgca acgagtcctg
     2881 tcccacagac acctacggac aaggatgcgc ccagcggtgc cgctgcgtgc accacaaggt
     2941 ttgccgcaag gccgacggta tgtgcatctg cgaaaccggc tggtcgggca cccggtgcga
     3001 tgaggtctgt ccggagggat tctacggcga gcactgcatg aatacctgtg cctgtccgtc
     3061 ggccaatttc cagtgccatg cggcccacgg ttgtgtgtgc aggagtggct acaccggcga
     3121 taactgcgac gagctaatcg cctctcaacg tattgcagac caaagcgaaa attcgagccg
     3181 ggcgagtgta gctctgacct tagtcctgat gaccctgttt gcgtacataa tcttcgcagt
     3241 gttcatttac taccgccgac gtgtgtccaa tctaaaaacc gaaatcgccc atgtacacta
     3301 cacccatgat acgaatcccc cgagctggcc gcccaaccac aacttcgaca atcccgtgta
     3361 cggcatgcag gcggaaacgc gtctcctgcc caacaatatg cgttccaaga tgaacaactt
     3421 tgaccagagg agcacgatga gcacggacta cggcgatgac tgcaatgcca gtggcagggt
     3481 gggtagctat tcgatcaact acaaccacga cctgctaacg aagaacttga acgcggaccg
     3541 gaccaatccg attgtgtaca acgaatccct caaagaggag catgtctacg acgagatcaa
     3601 gcacaaggag ggctacaagg atcccgatga atacgatcac ctggactact cacgacccag
     3661 cacctcgcag aagccgcact accaccgcat gaacgacgcc atgctcaaca tcaaccagga
     3721 cgaggagaag cccagcaacg tgaagaacat gactgtgctg ctgaacaaac cgctgccgcc
     3781 cacggaaccg gagccacaac acgagtgctt cgacaacacg aacaccaacc tggacaacgt
     3841 gtcgacggcg tcgcctagtt ccagtccgaa atttctaaag cggcgcgccg gcggccgctc
     3901 gagtctagag ggccccgtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     3961 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     4021 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     4081 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     4141 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     4201 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     4261 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     4321 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     4381 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     4441 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     4501 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     4561 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     4621 gtaagtttaa acccgctgat cagcctcgac tgtgccttct aagatccaga catgataaga
     4681 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     4741 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     4801 tctagaggat ccggtgcgtg gtggtttcat gcttctggga acggcaaatg ggtttaggat
     4861 tgggaacccc tcatcatctg ttggaatata ctattcaacc tacaaaagta acgttaaaca
     4921 acactacttt atatttgata tgaatggcca caccttttat gccataaaac atattgtaag
     4981 agaataccac tctttttatt ccttctttcc ttcttgtacg ttttttgctg taagtaggtc
     5041 gtggtgctgg tgttgcagtt gaaataactt aaaatataaa tcataaaact caaacataaa
     5101 cttgactatt tatttattta ttaagaaagg aaatataaat tataaattac aacagatccc
     5161 ccgggctgca ggaattcgat agcttgcatg cctgcaggtc ggccgccacg accggtgccg
     5221 ccaccatccc ctgacccacg cccctgaccc ctcacaagga gacgaccttc catgaccgag
     5281 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     5341 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     5401 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5461 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5521 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5581 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5641 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5701 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5761 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5821 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     5881 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atggctccga ccgaagccac
     5941 ccggggcggc cccgccgacc ccgcacccgc ccccgaggcc caccgactct agatcaagct
     6001 tatcgatacc gtcgacctcg aggccagcta gcttgaactt gtttattgca gcttataatg
     6061 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6121 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gggtacccaa
     6181 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     6241 aaacctctac aaatgtggta tggctgatta tgatcagtcg acctgcaggc atgcaagctt
     6301 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6361 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6421 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6481 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6541 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6601 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6661 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6721 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6781 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6841 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6901 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6961 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7021 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7081 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7141 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7201 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7261 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7321 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7381 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7441 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7501 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7561 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7621 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7681 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7741 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7801 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7861 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7921 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7981 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8041 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8101 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8161 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8221 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8281 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8341 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8461 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     8521 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     8581 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     8641 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
//
