LOCUS       sequence-251713-        8612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Draper-ITAM only-GFP mutant in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8612)
  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 8612)
  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 Draper-ITAM only-GFP
FEATURES             Location/Qualifiers
     source          1..8612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     91..110
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     304..326
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     318..335
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     319..335
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        341..767
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3928..4638
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVCGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3955..3983)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4578..4598
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4662..4679)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4681..4815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5290..5889
                     /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(5290..5309)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5786..5806
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6022..6038)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6023..6039)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6054..6188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6331..6347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6331..6347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6344..6366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6355..6371
                     /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(6379..6385,6386..6403,6404..6409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6424..6445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6562..6579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6733..7321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6813..6832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7492..8283,8284..8352))
                     /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     8115..8134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8353..8457)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8525..8543
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8581..8603)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
       61 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      121 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      181 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      241 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      301 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt aattcgttgc aggacaggat
      361 gtggtgcccg atgtgactag ctctttgctg caggccgtcc tatcctctgg ttccgataag
      421 agacccagaa ctccggcccc ccaccgccca ccgccacccc catacatatg tggtacgcaa
      481 gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag ctttgcatcc catacaagtc
      541 cccaaagtgg agaaccgaac caattcttcg cgggcagaac aaaagcttct gcacacgtct
      601 ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt gaatcgaacg aaagacccgt
      661 gtgtaaagcc gcgtttccaa aatgtataaa accgagagca tctggccaat gtgcatcagt
      721 tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa ctaaaggggg gatctagatc
      781 ggggtaccta ctagtcaaat gttgccggta atcctcatag cgtgcctggc ccagctggta
      841 ctggctcagg cggatcttaa agatctggat ggacccaata tctgcaaaag aagagaacta
      901 tataacgtag acgttgtcta cacagaactg caatcgttcc aggaacgggg ctccacctgg
      961 tgcgtcacat tcccgccgag gtgctccacc tatcgcatta aacaccgagt ggtcaacaaa
     1021 accaagacga ttgcgaagaa ccgtattgtg agggactgtt gtgatggcta catcgcgagc
     1081 gcaggagaat gcgtgccgca ttgttcggaa ccctgccagc acggtagatg catctccccg
     1141 gagaagtgca agtgtgatca tggttacgga ggacccgcct gcgatataaa ttgcccaccc
     1201 ggctggtatg gccggaactg ctcgatgcag tgcgactgcc tgaacaacgc ggtatgtgag
     1261 cccttttcag gcgactgcga gtgcgccaag ggctacactg gagcacgctg cgcggacatc
     1321 tgtccggagg gctttttcgg ggccaactgc tcggagaagt gccgatgcga gaacggcggc
     1381 aagtgtcacc acgtgtccgg cgaatgccag tgtgctcccg gtttcacggg tcccctatgt
     1441 gatatgcgct gtccggatgg aaagcatggc gcccagtgcc agcaggactg tccctgccag
     1501 aacgacggaa agtgtcagcc agagaccggg gcctgtatgt gcaatccagg ctggacgggc
     1561 gacgtgtgtg caaacaagtg tccggtgggc agctacggcc ctggatgcca agagagctgc
     1621 gagtgctaca agggagcgcc ctgccaccac atcaccggcc agtgcgagtg tccgcccgga
     1681 tatcgcggag aacgctgctt cgacgagtgt cagctgaaca cgtacggatt caactgcagc
     1741 atgacctgcg actgcgcgaa cgatgccatg tgcgatcggg cgaacggaac gtgcatctgc
     1801 aaccccggct ggacgggtgc caagtgcgcc gagcggatct gcgaggcgaa caagtacggg
     1861 ctggactgta accgcacctg cgagtgcgac atggagcaca cggatctgtg tcacccggag
     1921 acgggaaatt gccagtgcag catcggatgg agcagtgctc agtgcaccag accctgcacc
     1981 ttcctgcgct acggcccgaa ttgcgagctg acctgcaact gcaagaacgg agccaagtgc
     2041 tcgccggtca atggtacgtg cctgtgtgcg ccgggctggc gcggacccac ctgcgaggag
     2101 agctgcgagc cgggcacctt tggtcaggac tgcgccctgc gctgcgactg ccagaacgga
     2161 gccaagtgcg agccggagac gggccagtgc ttgtgcacgg ccggatggaa gaacatcaag
     2221 tgcgaccggc cgtgtgatct gaaccacttc ggacaggact gcgccaaggt gtgcgactgc
     2281 cacaataacg ccgcctgcaa tccgcagaac ggaagctgca cctgtgccgc cggctggacg
     2341 ggcgagcggt gcgaaaggaa gtgcgatacc ggcaaattcg gacacgactg cgcccagaag
     2401 tgccagtgcg acttcaacaa cagcctggcc tgcgatgcca ccaatggaag atgcgtgtgc
     2461 aagcaggatt ggggaggcgt tcattgcgag accaactgtc ggagcggtta ctatggtgaa
     2521 aattgtgata aagtttgcag atgcctgaat aactcctcct gcgaccccga ttccggtaac
     2581 tgcatctgct cagccggttg gactggcgcc gactgtgcgg aaccctgccc acctggattc
     2641 tacggcatgg aatgcaagga acggtgcccg gagatcctcc atggcaacaa gagctgcgat
     2701 cacatcacag gtgaaattct ctgccgcacg ggttacatag gtctcacctg cgagcaccca
     2761 tgtcctgctg gactctacgg acccggctgc aagctcaagt gcaattgcga gcacggcggc
     2821 gagtgcaacc acgttacagg ccagtgccag tgcctgccag ggtggacagg ttccaactgc
     2881 aacgagtcct gtcccacaga cacctacgga caaggatgcg cccagcggtg ccgctgcgtg
     2941 caccacaagg tttgccgcaa ggccgacggt atgtgcatct gcgaaaccgg ctggtcgggc
     3001 acccggtgcg atgaggtctg tccggaggga ttctacggcg agcactgcat gaatacctgt
     3061 gcctgtccgt cggccaattt ccagtgccat gcggcccacg gttgtgtgtg caggagtggc
     3121 tacaccggcg ataactgcga cgagctaatc gcctctcaac gtattgcaga ccaaagcgaa
     3181 aattcgagcc gggcgagtgt agctctgacc ttagtcctga tgaccctgtt tgcgtgcata
     3241 atcttcgcag tgttcattta ctaccgccga cgtgtgtcca atctaaaaac cgaaatcgcc
     3301 catgtacact tcacccatga tacgaatccc ccgagctggc cgcccaacca caacttcgac
     3361 aatcccgtgt tcggcatgca ggcggaaacg cgtctcctgc ccaacaatat gcgttccaag
     3421 atgaacaact ttgaccagag gagcacgatg agcacggact tcggcgatga ctgcaatgcc
     3481 agtggcaggg tgggtagctt ctcgatcaac ttcaaccacg acctgctaac gaagaacttg
     3541 aacgcggacc ggaccaatcc gattgtgttc aacgaatccc tcaaagagga gcatgtctac
     3601 gacgagatca agcacaagga gggcttcaag gatcccgatg aatacgatca cctggacttc
     3661 tcacgaccca gcacctcgca gaagccgcac ttccaccgca tgaacgacgc catgctcaac
     3721 atcaaccagg acgaggagaa gcccagcaac gtgaagaaca tgactgtgct gctgaacaaa
     3781 ccgctgccgc ccacggaacc ggagccacaa cacgagtgct tcgacaacac gaacaccaac
     3841 ctggacaacg tgtcgacggc gtcgcctagt tccagtccga aatttctaaa gcggcgcgcc
     3901 ggcggccgct cgagtctaga gggccccagc aaaggagaag aacttttcac tggagttgtc
     3961 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt ctgtggagag
     4021 ggtgaaggtg atgctacaaa cggaaaactc acccttaaat ttatttgcac tactggaaaa
     4081 ctacctgttc cgtggccaac acttgtcact actctgacct atggtgttca atgcttttcc
     4141 cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     4201 gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg tgctgaagtc
     4261 aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga ttttaaagaa
     4321 gatggaaaca ttcttggaca caaactcgag tacaacttta actcacacaa tgtatacatc
     4381 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacgttgaa
     4441 gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     4501 gtccttttac cagacaacca ttacctgtcg acacaatctg tcctttcgaa agatcccaac
     4561 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
     4621 atggatgagc tctacaaata gtttaaacgc ccgctgatca gcctcgactg tgccttctaa
     4681 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4741 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4801 tgcaataaac aagttaactc tagaggatcc ggtgcgtggt ggtttcatgc ttctgggaac
     4861 ggcaaatggg tttaggattg ggaacccctc atcatctgtt ggaatatact attcaaccta
     4921 caaaagtaac gttaaacaac actactttat atttgatatg aatggccaca ccttttatgc
     4981 cataaaacat attgtaagag aataccactc tttttattcc ttctttcctt cttgtacgtt
     5041 ttttgctgta agtaggtcgt ggtgctggtg ttgcagttga aataacttaa aatataaatc
     5101 ataaaactca aacataaact tgactattta tttatttatt aagaaaggaa atataaatta
     5161 taaattacaa cagatccccc gggctgcagg aattcgatag cttgcatgcc tgcaggtcgg
     5221 ccgccacgac cggtgccgcc accatcccct gacccacgcc cctgacccct cacaaggaga
     5281 cgaccttcca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5341 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5401 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5461 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5521 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     5581 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5641 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5701 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5761 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5821 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5881 ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat
     5941 ggctccgacc gaagccaccc ggggcggccc cgccgacccc gcacccgccc ccgaggccca
     6001 ccgactctag atcaagctta tcgataccgt cgacctcgag gccagctagc ttgaacttgt
     6061 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6121 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6181 tctggatcgg gtacccaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     6241 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     6301 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6361 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6421 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6481 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6541 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6661 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6901 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7141 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7381 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7441 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7501 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7561 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7621 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7681 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7741 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7801 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7861 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7921 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7981 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8041 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8101 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8161 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8221 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8281 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8341 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8401 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8461 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     8521 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     8581 cctctgacac atgcagctcc cggagacggt ca
//
