LOCUS       sequence-251709-        8613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Draper-GFP in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8613)
  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 8613)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..8613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     189..211
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     203..220
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     204..220
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        226..652
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3813..4526
                     /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             3813..4523
                     /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(3840..3868)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4463..4483
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4548..4565)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4567..4701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5176..5775
                     /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(5176..5195)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5672..5692
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5908..5924)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5909..5925)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5940..6074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6217..6233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6217..6233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6230..6252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6241..6257
                     /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(6265..6271,6272..6289,6290..6295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6310..6331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6448..6465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6619..7207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6699..6718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7378..8169,8170..8238))
                     /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     8001..8020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8239..8343)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8411..8429
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8467..8489)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8589..8608
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
       61 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      121 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      181 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattaattc gttgcaggac
      241 aggatgtggt gcccgatgtg actagctctt tgctgcaggc cgtcctatcc tctggttccg
      301 ataagagacc cagaactccg gccccccacc gcccaccgcc acccccatac atatgtggta
      361 cgcaagtaag agtgcctgcg catgccccat gtgccccacc aagagctttg catcccatac
      421 aagtccccaa agtggagaac cgaaccaatt cttcgcgggc agaacaaaag cttctgcaca
      481 cgtctccact cgaatttgga gccggccggc gtgtgcaaaa gaggtgaatc gaacgaaaga
      541 cccgtgtgta aagccgcgtt tccaaaatgt ataaaaccga gagcatctgg ccaatgtgca
      601 tcagttgtgg tcagcagcaa aatcaagtga atcatctcag tgcaactaaa ggggggatct
      661 agatcggggt acctactagt caaatgttgc cggtaatcct catagcgtgc ctggcccagc
      721 tggtactggc tcaggcggat cttaaagatc tggatggacc caatatctgc aaaagaagag
      781 aactatataa cgtagacgtt gtctacacag aactgcaatc gttccaggaa cggggctcca
      841 cctggtgcgt cacattcccg ccgaggtgct ccacctatcg cattaaacac cgagtggtca
      901 acaaaaccaa gacgattgcg aagaaccgta ttgtgaggga ctgttgtgat ggctacatcg
      961 cgagcgcagg agaatgcgtg ccgcattgtt cggaaccctg ccagcacggt agatgcatct
     1021 ccccggagaa gtgcaagtgt gatcatggtt acggaggacc cgcctgcgat ataaattgcc
     1081 cacccggctg gtatggccgg aactgctcga tgcagtgcga ctgcctgaac aacgcggtat
     1141 gtgagccctt ttcaggcgac tgcgagtgcg ccaagggcta cactggagca cgctgcgcgg
     1201 acatctgtcc ggagggcttt ttcggggcca actgctcgga gaagtgccga tgcgagaacg
     1261 gcggcaagtg tcaccacgtg tccggcgaat gccagtgtgc tcccggtttc acgggtcccc
     1321 tatgtgatat gcgctgtccg gatggaaagc atggcgccca gtgccagcag gactgtccct
     1381 gccagaacga cggaaagtgt cagccagaga ccggggcctg tatgtgcaat ccaggctgga
     1441 cgggcgacgt gtgtgcaaac aagtgtccgg tgggcagcta cggccctgga tgccaagaga
     1501 gctgcgagtg ctacaaggga gcgccctgcc accacatcac cggccagtgc gagtgtccgc
     1561 ccggatatcg cggagaacgc tgcttcgacg agtgtcagct gaacacgtac ggattcaact
     1621 gcagcatgac ctgcgactgc gcgaacgatg ccatgtgcga tcgggcgaac ggaacgtgca
     1681 tctgcaaccc cggctggacg ggtgccaagt gcgccgagcg gatctgcgag gcgaacaagt
     1741 acgggctgga ctgtaaccgc acctgcgagt gcgacatgga gcacacggat ctgtgtcacc
     1801 cggagacggg aaattgccag tgcagcatcg gatggagcag tgctcagtgc accagaccct
     1861 gcaccttcct gcgctacggc ccgaattgcg agctgacctg caactgcaag aacggagcca
     1921 agtgctcgcc ggtcaatggt acgtgcctgt gtgcgccggg ctggcgcgga cccacctgcg
     1981 aggagagctg cgagccgggc acctttggtc aggactgcgc cctgcgctgc gactgccaga
     2041 acggagccaa gtgcgagccg gagacgggcc agtgcttgtg cacggccgga tggaagaaca
     2101 tcaagtgcga ccggccgtgt gatctgaacc acttcggaca ggactgcgcc aaggtgtgcg
     2161 actgccacaa taacgccgcc tgcaatccgc agaacggaag ctgcacctgt gccgccggct
     2221 ggacgggcga gcggtgcgaa aggaagtgcg ataccggcaa attcggacac gactgcgccc
     2281 agaagtgcca gtgcgacttc aacaacagcc tggcctgcga tgccaccaat ggaagatgcg
     2341 tgtgcaagca ggattgggga ggcgttcatt gcgagaccaa ctgtcggagc ggttactatg
     2401 gtgaaaattg tgataaagtt tgcagatgcc tgaataactc ctcctgcgac cccgattccg
     2461 gtaactgcat ctgctcagcc ggttggactg gcgccgactg tgcggaaccc tgcccacctg
     2521 gattctacgg catggaatgc aaggaacggt gcccggagat cctccatggc aacaagagct
     2581 gcgatcacat cacaggtgaa attctctgcc gcacgggtta cataggtctc acctgcgagc
     2641 acccatgtcc tgctggactc tacggacccg gctgcaagct caagtgcaat tgcgagcacg
     2701 gcggcgagtg caaccacgtt acaggccagt gccagtgcct gccagggtgg acaggttcca
     2761 actgcaacga gtcctgtccc acagacacct acggacaagg atgcgcccag cggtgccgct
     2821 gcgtgcacca caaggtttgc cgcaaggccg acggtatgtg catctgcgaa accggctggt
     2881 cgggcacccg gtgcgatgag gtctgtccgg agggattcta cggcgagcac tgcatgaata
     2941 cctgtgcctg tccgtcggcc aatttccagt gccatgcggc ccacggttgt gtgtgcagga
     3001 gtggctacac cggcgataac tgcgacgagc taatcgcctc tcaacgtatt gcagaccaaa
     3061 gcgaaaattc gagccgggcg agtgtagctc tgaccttagt cctgatgacc ctgtttgcgt
     3121 gcataatctt cgcagtgttc atttactacc gccgacgtgt gtccaatcta aaaaccgaaa
     3181 tcgcccatgt acactacacc catgatacga atcccccgag ctggccgccc aaccacaact
     3241 tcgacaatcc cgtgtacggc atgcaggcgg aaacgcgtct cctgcccaac aatatgcgtt
     3301 ccaagatgaa caactttgac cagaggagca cgatgagcac ggactacggc gatgactgca
     3361 atgccagtgg cagggtgggt agctattcga tcaactacaa ccacgacctg ctaacgaaga
     3421 acttgaacgc ggaccggacc aatccgattg tgtacaacga atccctcaaa gaggagcatg
     3481 tctacgacga gatcaagcac aaggagggct acaaggatcc cgatgaatac gatcacctgg
     3541 actactcacg acccagcacc tcgcagaagc cgcactacca ccgcatgaac gacgccatgc
     3601 tcaacatcaa ccaggacgag gagaagccca gcaacgtgaa gaacatgact gtgctgctga
     3661 acaaaccgct gccgcccacg gaaccggagc cacaacacga gtgcttcgac aacacgaaca
     3721 ccaacctgga caacgtgtcg acggcgtcgc ctagttccag tccgaaattt ctaaagcggc
     3781 gcgccggcgg ccgctcgagt ctagagggcc ccagcaaagg agaagaactt ttcactggag
     3841 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtccgtg
     3901 gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt tgcactactg
     3961 gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt gttcaatgct
     4021 tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc atgcccgaag
     4081 gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag acgcgtgctg
     4141 aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt attgatttta
     4201 aagaagatgg aaacattctt ggacacaaac tcgagtacaa ctttaactca cacaatgtat
     4261 acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaacg
     4321 ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     4381 gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt tcgaaagatc
     4441 ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac
     4501 atggcatgga tgagctctac aaataggttt aaacgcccgc tgatcagcct cgactgtgcc
     4561 ttctaagatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4621 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4681 ataagctgca ataaacaagt taactctaga ggatccggtg cgtggtggtt tcatgcttct
     4741 gggaacggca aatgggttta ggattgggaa cccctcatca tctgttggaa tatactattc
     4801 aacctacaaa agtaacgtta aacaacacta ctttatattt gatatgaatg gccacacctt
     4861 ttatgccata aaacatattg taagagaata ccactctttt tattccttct ttccttcttg
     4921 tacgtttttt gctgtaagta ggtcgtggtg ctggtgttgc agttgaaata acttaaaata
     4981 taaatcataa aactcaaaca taaacttgac tatttattta tttattaaga aaggaaatat
     5041 aaattataaa ttacaacaga tcccccgggc tgcaggaatt cgatagcttg catgcctgca
     5101 ggtcggccgc cacgaccggt gccgccacca tcccctgacc cacgcccctg acccctcaca
     5161 aggagacgac cttccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     5221 gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     5281 accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     5341 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5401 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5461 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     5521 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5581 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5641 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     5701 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     5761 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     5821 ccccatggct ccgaccgaag ccacccgggg cggccccgcc gaccccgcac ccgcccccga
     5881 ggcccaccga ctctagatca agcttatcga taccgtcgac ctcgaggcca gctagcttga
     5941 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6001 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6061 atcatgtctg gatcgggtac ccaattgcat tcattttatg tttcaggttc agggggaggt
     6121 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca
     6181 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6241 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6301 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6361 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6421 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6481 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6541 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7021 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8341 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8401 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     8461 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     8521 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     8581 taactatgcg gcatcagagc agattgtact gag
//
