LOCUS       sequence-251710-        8612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Draper-Ala11-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-Ala11-GFP
FEATURES             Location/Qualifiers
     source          1..8612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     198..220
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     212..229
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     213..229
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        235..661
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3822..4535
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             3822..4532
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3849..3877)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4472..4492
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4556..4573)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4575..4709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5184..5783
                     /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(5184..5203)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5680..5700
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5916..5932)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5917..5933)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5948..6082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6225..6241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6225..6241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6238..6260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6249..6265
                     /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(6273..6279,6280..6297,6298..6303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6318..6339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6456..6473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6627..7215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6707..6726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7386..8177,8178..8246))
                     /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     8009..8028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8247..8351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8419..8437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8475..8497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     join(8597..8612,1..4)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
       61 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      121 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      181 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattaattcg
      241 ttgcaggaca ggatgtggtg cccgatgtga ctagctcttt gctgcaggcc gtcctatcct
      301 ctggttccga taagagaccc agaactccgg ccccccaccg cccaccgcca cccccataca
      361 tatgtggtac gcaagtaaga gtgcctgcgc atgccccatg tgccccacca agagctttgc
      421 atcccataca agtccccaaa gtggagaacc gaaccaattc ttcgcgggca gaacaaaagc
      481 ttctgcacac gtctccactc gaatttggag ccggccggcg tgtgcaaaag aggtgaatcg
      541 aacgaaagac ccgtgtgtaa agccgcgttt ccaaaatgta taaaaccgag agcatctggc
      601 caatgtgcat cagttgtggt cagcagcaaa atcaagtgaa tcatctcagt gcaactaaag
      661 gggggatcta gatcggggta cctactagtc aaatgttgcc ggtaatcctc atagcgtgcc
      721 tggcccagct ggtactggct caggcggatc ttaaagatct ggatggaccc aatatctgca
      781 aaagaagaga actatataac gtagacgttg tctacacaga actgcaatcg ttccaggaac
      841 ggggctccac ctggtgcgtc acattcccgc cgaggtgctc cacctatcgc attaaacacc
      901 gagtggtcaa caaaaccaag acgattgcga agaaccgtat tgtgagggac tgttgtgatg
      961 gctacatcgc gagcgcagga gaatgcgtgc cgcattgttc ggaaccctgc cagcacggta
     1021 gatgcatctc cccggagaag tgcaagtgtg atcatggtta cggaggaccc gcctgcgata
     1081 taaattgccc acccggctgg tatggccgga actgctcgat gcagtgcgac tgcctgaaca
     1141 acgcggtatg tgagcccttt tcaggcgact gcgagtgcgc caagggctac actggagcac
     1201 gctgcgcgga catctgtccg gagggctttt tcggggccaa ctgctcggag aagtgccgat
     1261 gcgagaacgg cggcaagtgt caccacgtgt ccggcgaatg ccagtgtgct cccggtttca
     1321 cgggtcccct atgtgatatg cgctgtccgg atggaaagca tggcgcccag tgccagcagg
     1381 actgtccctg ccagaacgac ggaaagtgtc agccagagac cggggcctgt atgtgcaatc
     1441 caggctggac gggcgacgtg tgtgcaaaca agtgtccggt gggcagctac ggccctggat
     1501 gccaagagag ctgcgagtgc tacaagggag cgccctgcca ccacatcacc ggccagtgcg
     1561 agtgtccgcc cggatatcgc ggagaacgct gcttcgacga gtgtcagctg aacacgtacg
     1621 gattcaactg cagcatgacc tgcgactgcg cgaacgatgc catgtgcgat cgggcgaacg
     1681 gaacgtgcat ctgcaacccc ggctggacgg gtgccaagtg cgccgagcgg atctgcgagg
     1741 cgaacaagta cgggctggac tgtaaccgca cctgcgagtg cgacatggag cacacggatc
     1801 tgtgtcaccc ggagacggga aattgccagt gcagcatcgg atggagcagt gctcagtgca
     1861 ccagaccctg caccttcctg cgctacggcc cgaattgcga gctgacctgc aactgcaaga
     1921 acggagccaa gtgctcgccg gtcaatggta cgtgcctgtg tgcgccgggc tggcgcggac
     1981 ccacctgcga ggagagctgc gagccgggca cctttggtca ggactgcgcc ctgcgctgcg
     2041 actgccagaa cggagccaag tgcgagccgg agacgggcca gtgcttgtgc acggccggat
     2101 ggaagaacat caagtgcgac cggccgtgtg atctgaacca cttcggacag gactgcgcca
     2161 aggtgtgcga ctgccacaat aacgccgcct gcaatccgca gaacggaagc tgcacctgtg
     2221 ccgccggctg gacgggcgag cggtgcgaaa ggaagtgcga taccggcaaa ttcggacacg
     2281 actgcgccca gaagtgccag tgcgacttca acaacagcct ggcctgcgat gccaccaatg
     2341 gaagatgcgt gtgcaagcag gattggggag gcgttcattg cgagaccaac tgtcggagcg
     2401 gttactatgg tgaaaattgt gataaagttt gcagatgcct gaataactcc tcctgcgacc
     2461 ccgattccgg taactgcatc tgctcagccg gttggactgg cgccgactgt gcggaaccct
     2521 gcccacctgg attctacggc atggaatgca aggaacggtg cccggagatc ctccatggca
     2581 acaagagctg cgatcacatc acaggtgaaa ttctctgccg cacgggttac ataggtctca
     2641 cctgcgagca cccatgtcct gctggactct acggacccgg ctgcaagctc aagtgcaatt
     2701 gcgagcacgg cggcgagtgc aaccacgtta caggccagtg ccagtgcctg ccagggtgga
     2761 caggttccaa ctgcaacgag tcctgtccca cagacaccta cggacaagga tgcgcccagc
     2821 ggtgccgctg cgtgcaccac aaggtttgcc gcaaggccga cggtatgtgc atctgcgaaa
     2881 ccggctggtc gggcacccgg tgcgatgagg tctgtccgga gggattctac ggcgagcact
     2941 gcatgaatac ctgtgcctgt ccgtcggcca atttccagtg ccatgcggcc cacggttgtg
     3001 tgtgcaggag tggctacacc ggcgataact gcgacgagct aatcgcctct caacgtattg
     3061 cagaccaaag cgaaaattcg agccgggcga gtgtagctct gaccttagtc ctgatgaccc
     3121 tgtttgcgtg cataatcttc gcagtgttca tttactaccg ccgacgtgtg tccaatctaa
     3181 aaaccgaaat cgcccatgta cacgcaaccc atgatacgaa tcccccgagc tggccgccca
     3241 accacaactt cgacaatccc gtggcaggca tgcaggcgga aacgcgtctc ctgcccaaca
     3301 atatgcgttc caagatgaac aactttgacc agaggagcac gatgagcacg gacgcaggcg
     3361 atgactgcaa tgccagtggc agggtgggta gcgcatcgat caacgcaaac cacgacctgc
     3421 taacgaagaa cttgaacgcg gaccggacca atccgattgt ggcaaacgaa tccctcaaag
     3481 aggagcatgt cgcagacgag atcaagcaca aggagggcgc aaaggatccc gatgaagcag
     3541 atcacctgga cgcatcacga cccagcacct cgcagaagcc gcacgcacac cgcatgaacg
     3601 acgccatgct caacatcaac caggacgagg agaagcccag caacgtgaag aacatgactg
     3661 tgctgctgaa caaaccgctg ccgcccacgg aaccggagcc acaacacgag tgcttcgaca
     3721 acacgaacac caacctggac aacgtgtcga cggcgtcgcc tagttccagt ccgaaatttc
     3781 taaagcggcg cgccggcggc cgctcgagtc tagagggccc cagcaaagga gaagaacttt
     3841 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     3901 ctgtccgtgg agagggtgaa ggtgatgcta caaacggaaa actcaccctt aaatttattt
     3961 gcactactgg aaaactacct gttccgtggc caacacttgt cactactctg acctatggtg
     4021 ttcaatgctt ttcccgttat ccggatcaca tgaaacggca tgactttttc aagagtgcca
     4081 tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg acctacaaga
     4141 cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag ttaaagggta
     4201 ttgattttaa agaagatgga aacattcttg gacacaaact cgagtacaac tttaactcac
     4261 acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaattc
     4321 gccacaacgt tgaagatggt tccgttcaac tagcagacca ttatcaacaa aatactccaa
     4381 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa tctgtccttt
     4441 cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta actgctgctg
     4501 ggattacaca tggcatggat gagctctaca aataggttaa acgcccgctg atcagcctcg
     4561 actgtgcctt ctaagatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     4621 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4681 taaccattat aagctgcaat aaacaagtta actctagagg atccggtgcg tggtggtttc
     4741 atgcttctgg gaacggcaaa tgggtttagg attgggaacc cctcatcatc tgttggaata
     4801 tactattcaa cctacaaaag taacgttaaa caacactact ttatatttga tatgaatggc
     4861 cacacctttt atgccataaa acatattgta agagaatacc actcttttta ttccttcttt
     4921 ccttcttgta cgttttttgc tgtaagtagg tcgtggtgct ggtgttgcag ttgaaataac
     4981 ttaaaatata aatcataaaa ctcaaacata aacttgacta tttatttatt tattaagaaa
     5041 ggaaatataa attataaatt acaacagatc ccccgggctg caggaattcg atagcttgca
     5101 tgcctgcagg tcggccgcca cgaccggtgc cgccaccatc ccctgaccca cgcccctgac
     5161 ccctcacaag gagacgacct tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     5221 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     5281 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     5341 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     5401 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     5461 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     5521 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     5581 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5641 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     5701 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5761 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
     5821 gcgcacgacc ccatggctcc gaccgaagcc acccggggcg gccccgccga ccccgcaccc
     5881 gcccccgagg cccaccgact ctagatcaag cttatcgata ccgtcgacct cgaggccagc
     5941 tagcttgaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6001 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6061 tgtatcttat catgtctgga tcgggtaccc aattgcattc attttatgtt tcaggttcag
     6121 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     6181 tatgatcagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
     6241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8401 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     8461 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     8521 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     8581 ggctggctta actatgcggc atcagagcag at
//
