LOCUS       sequence-251711-        8613 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Inducible expression of Draper-delta ITAM-GFP mutant 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 Apr 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-delta ITAM-GFP
FEATURES             Location/Qualifiers
     source          1..8613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..80
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     72..89
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     73..89
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        95..521
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3682..4395
                     /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             3682..4392
                     /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(3709..3737)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4332..4352
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4417..4434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4436..4570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5045..5644
                     /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(5045..5064)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5541..5561
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5777..5793)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5778..5794)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5809..5943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6086..6102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6086..6102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6099..6121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6110..6126
                     /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(6134..6140,6141..6158,6159..6164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6179..6200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6317..6334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6488..7076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6568..6587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7247..8038,8039..8107))
                     /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     7870..7889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8108..8212)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8280..8298
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8336..8358)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8458..8477
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
       61 agtcacgacg ttgtaaaacg acggccagtg aattaattcg ttgcaggaca ggatgtggtg
      121 cccgatgtga ctagctcttt gctgcaggcc gtcctatcct ctggttccga taagagaccc
      181 agaactccgg ccccccaccg cccaccgcca cccccataca tatgtggtac gcaagtaaga
      241 gtgcctgcgc atgccccatg tgccccacca agagctttgc atcccataca agtccccaaa
      301 gtggagaacc gaaccaattc ttcgcgggca gaacaaaagc ttctgcacac gtctccactc
      361 gaatttggag ccggccggcg tgtgcaaaag aggtgaatcg aacgaaagac ccgtgtgtaa
      421 agccgcgttt ccaaaatgta taaaaccgag agcatctggc caatgtgcat cagttgtggt
      481 cagcagcaaa atcaagtgaa tcatctcagt gcaactaaag gggggatcta gatcggggta
      541 cctactagtc aaatgttgcc ggtaatcctc atagcgtgcc tggcccagct ggtactggct
      601 caggcggatc ttaaagatct ggatggaccc aatatctgca aaagaagaga actatataac
      661 gtagacgttg tctacacaga actgcaatcg ttccaggaac ggggctccac ctggtgcgtc
      721 acattcccgc cgaggtgctc cacctatcgc attaaacacc gagtggtcaa caaaaccaag
      781 acgattgcga agaaccgtat tgtgagggac tgttgtgatg gctacatcgc gagcgcagga
      841 gaatgcgtgc cgcattgttc ggaaccctgc cagcacggta gatgcatctc cccggagaag
      901 tgcaagtgtg atcatggtta cggaggaccc gcctgcgata taaattgccc acccggctgg
      961 tatggccgga actgctcgat gcagtgcgac tgcctgaaca acgcggtatg tgagcccttt
     1021 tcaggcgact gcgagtgcgc caagggctac actggagcac gctgcgcgga catctgtccg
     1081 gagggctttt tcggggccaa ctgctcggag aagtgccgat gcgagaacgg cggcaagtgt
     1141 caccacgtgt ccggcgaatg ccagtgtgct cccggtttca cgggtcccct atgtgatatg
     1201 cgctgtccgg atggaaagca tggcgcccag tgccagcagg actgtccctg ccagaacgac
     1261 ggaaagtgtc agccagagac cggggcctgt atgtgcaatc caggctggac gggcgacgtg
     1321 tgtgcaaaca agtgtccggt gggcagctac ggccctggat gccaagagag ctgcgagtgc
     1381 tacaagggag cgccctgcca ccacatcacc ggccagtgcg agtgtccgcc cggatatcgc
     1441 ggagaacgct gcttcgacga gtgtcagctg aacacgtacg gattcaactg cagcatgacc
     1501 tgcgactgcg cgaacgatgc catgtgcgat cgggcgaacg gaacgtgcat ctgcaacccc
     1561 ggctggacgg gtgccaagtg cgccgagcgg atctgcgagg cgaacaagta cgggctggac
     1621 tgtaaccgca cctgcgagtg cgacatggag cacacggatc tgtgtcaccc ggagacggga
     1681 aattgccagt gcagcatcgg atggagcagt gctcagtgca ccagaccctg caccttcctg
     1741 cgctacggcc cgaattgcga gctgacctgc aactgcaaga acggagccaa gtgctcgccg
     1801 gtcaatggta cgtgcctgtg tgcgccgggc tggcgcggac ccacctgcga ggagagctgc
     1861 gagccgggca cctttggtca ggactgcgcc ctgcgctgcg actgccagaa cggagccaag
     1921 tgcgagccgg agacgggcca gtgcttgtgc acggccggat ggaagaacat caagtgcgac
     1981 cggccgtgtg atctgaacca cttcggacag gactgcgcca aggtgtgcga ctgccacaat
     2041 aacgccgcct gcaatccgca gaacggaagc tgcacctgtg ccgccggctg gacgggcgag
     2101 cggtgcgaaa ggaagtgcga taccggcaaa ttcggacacg actgcgccca gaagtgccag
     2161 tgcgacttca acaacagcct ggcctgcgat gccaccaatg gaagatgcgt gtgcaagcag
     2221 gattggggag gcgttcattg cgagaccaac tgtcggagcg gttactatgg tgaaaattgt
     2281 gataaagttt gcagatgcct gaataactcc tcctgcgacc ccgattccgg taactgcatc
     2341 tgctcagccg gttggactgg cgccgactgt gcggaaccct gcccacctgg attctacggc
     2401 atggaatgca aggaacggtg cccggagatc ctccatggca acaagagctg cgatcacatc
     2461 acaggtgaaa ttctctgccg cacgggttac ataggtctca cctgcgagca cccatgtcct
     2521 gctggactct acggacccgg ctgcaagctc aagtgcaatt gcgagcacgg cggcgagtgc
     2581 aaccacgtta caggccagtg ccagtgcctg ccagggtgga caggttccaa ctgcaacgag
     2641 tcctgtccca cagacaccta cggacaagga tgcgcccagc ggtgccgctg cgtgcaccac
     2701 aaggtttgcc gcaaggccga cggtatgtgc atctgcgaaa ccggctggtc gggcacccgg
     2761 tgcgatgagg tctgtccgga gggattctac ggcgagcact gcatgaatac ctgtgcctgt
     2821 ccgtcggcca atttccagtg ccatgcggcc cacggttgtg tgtgcaggag tggctacacc
     2881 ggcgataact gcgacgagct aatcgcctct caacgtattg cagaccaaag cgaaaattcg
     2941 agccgggcga gtgtagctct gaccttagtc ctgatgaccc tgtttgcgtg cataatcttc
     3001 gcagtgttca tttactaccg ccgacgtgtg tccaatctaa aaaccgaaat cgcccatgta
     3061 cactacaccc atgatacgaa tcccccgagc tggccgccca accacaactt cgacaatccc
     3121 gtgtacggca tgcaggcgga aacgcgtctc ctgcccaaca atatgcgttc caagatgaac
     3181 aactttgacc agaggagcac gatgagcacg gactacggcg atgactgcaa tgccagtggc
     3241 agggtgggta gctattcgat caactacaac cacgacctgc taacgaagaa cttgaacgcg
     3301 gaccggacca atccgattgt gtacaacgaa tccctcaaag aggagcatgt cttcgacgag
     3361 atcaagcaca aggagggcta caaggatccc gatgaattcg atcacctgga ctactcacga
     3421 cccagcacct cgcagaagcc gcactaccac cgcatgaacg acgccatgct caacatcaac
     3481 caggacgagg agaagcccag caacgtgaag aacatgactg tgctgctgaa caaaccgctg
     3541 ccgcccacgg aaccggagcc acaacacgag tgcttcgaca acacgaacac caacctggac
     3601 aacgtgtcga cggcgtcgcc tagttccagt ccgaaatttc taaagcggcg cgccggcggc
     3661 cgctcgagtc tagagggccc cagcaaagga gaagaacttt tcactggagt tgtcccaatt
     3721 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa
     3781 ggtgatgcta caaacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
     3841 gttccgtggc caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat
     3901 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     3961 gaacgcacta tatctttcaa agatgacggg acctacaaga cgcgtgctga agtcaagttt
     4021 gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga
     4081 aacattcttg gacacaaact cgagtacaac tttaactcac acaatgtata catcacggca
     4141 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacgt tgaagatggt
     4201 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     4261 ttaccagaca accattacct gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag
     4321 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
     4381 gagctctaca aataggttta aacgcccgct gatcagcctc gactgtgcct tctaagatcc
     4441 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4501 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4561 taaacaagtt aactctagag gatccggtgc gtggtggttt catgcttctg ggaacggcaa
     4621 atgggtttag gattgggaac ccctcatcat ctgttggaat atactattca acctacaaaa
     4681 gtaacgttaa acaacactac tttatatttg atatgaatgg ccacaccttt tatgccataa
     4741 aacatattgt aagagaatac cactcttttt attccttctt tccttcttgt acgttttttg
     4801 ctgtaagtag gtcgtggtgc tggtgttgca gttgaaataa cttaaaatat aaatcataaa
     4861 actcaaacat aaacttgact atttatttat ttattaagaa aggaaatata aattataaat
     4921 tacaacagat cccccgggct gcaggaattc gatagcttgc atgcctgcag gtcggccgcc
     4981 acgaccggtg ccgccaccat cccctgaccc acgcccctga cccctcacaa ggagacgacc
     5041 ttccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     5101 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     5161 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     5221 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     5281 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5341 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5401 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5461 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5521 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5581 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5641 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc
     5701 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac
     5761 tctagatcaa gcttatcgat accgtcgacc tcgaggccag ctagcttgaa cttgtttatt
     5821 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5881 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5941 atcgggtacc caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     6001 tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgacctgca
     6061 ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     6121 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6181 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6241 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6301 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6361 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6421 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6481 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6661 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6841 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6901 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6961 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     7021 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7081 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7141 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7321 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7561 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7861 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7981 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     8041 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8101 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8161 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8221 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8281 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     8341 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     8401 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     8461 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     8521 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     8581 ggcgatcggt gcgggcctct tcgctattac gcc
//
