LOCUS       sequence-251675-        8301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of Shark-TagBFP in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8301)
  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 8301)
  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 Shark-TagBFP
FEATURES             Location/Qualifiers
     source          1..8301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..136
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     330..352
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     344..361
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     345..361
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        367..793
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3654..4355
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(4377..4394)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4396..4530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5005..5604
                     /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(5005..5024)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5501..5521
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5737..5753)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5738..5754)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5769..5903
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6046..6062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6046..6062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6059..6081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6070..6086
                     /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(6094..6100,6101..6118,6119..6124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6139..6160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6277..6294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6448..7036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6528..6547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7207..7998,7999..8067))
                     /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     7830..7849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8068..8172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8240..8258
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(8296..8301,1..17))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
       61 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
      121 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
      181 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
      241 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
      301 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      361 tgaattaatt cgttgcagga caggatgtgg tgcccgatgt gactagctct ttgctgcagg
      421 ccgtcctatc ctctggttcc gataagagac ccagaactcc ggccccccac cgcccaccgc
      481 cacccccata catatgtggt acgcaagtaa gagtgcctgc gcatgcccca tgtgccccac
      541 caagagcttt gcatcccata caagtcccca aagtggagaa ccgaaccaat tcttcgcggg
      601 cagaacaaaa gcttctgcac acgtctccac tcgaatttgg agccggccgg cgtgtgcaaa
      661 agaggtgaat cgaacgaaag acccgtgtgt aaagccgcgt ttccaaaatg tataaaaccg
      721 agagcatctg gccaatgtgc atcagttgtg gtcagcagca aaatcaagtg aatcatctca
      781 gtgcaactaa aggggggatc tagatcgggg tacctactag tcaaatgagt cgcgatagtg
      841 atcccatgaa atggtaccac ggcaacctgt cccgcgaggc agccgacgaa ctgctaaagc
      901 agggttacga ggacgggaca tttctggtgc gcgagagcag cacagcggcc ggggactttg
      961 tgctgagcct gctctgccag ggtgaggtgt gtcactacca ggtgcggcga catggcggcg
     1021 aggacgcctt cttctccatc gacgacaagg tacagacgaa gatcctgcac ggcctggaca
     1081 cgctcgtcga ctactatcag caggcggcca acggtctgcc caccaagctg acggtgccct
     1141 tgatccgcga tctgccgccg cacaacactc gcagccacgg agtgaccaat ctgctgcatc
     1201 gcgccacaag taaaaacgag agcaaggtcg tcttcgagct gctcaagtgc ggctatcgga
     1261 atttcgatgc caagaatcag gatggacaga cggctctgca tctggctgca ctccactcgg
     1321 acgaggatat actgaaacac ctgttgaatg caaaggtcca ggtcaacagc tccgactcct
     1381 ttggctgcca gccattgcac tacgccgctc gctccaaacc ggccagtttc atacgcacac
     1441 tcatctcggc acaggccaat gtccagggca ggaacatcga caacggctat gtgccgctgc
     1501 atgaggctgc caagcacggc aacctggagg ctgtgcaaga gctgctcctg gctgaggctc
     1561 cgccgttgcc cagaacaagt tcgggtgaat ttccattcga tctggccaag gaagcgggcc
     1621 aaacggcagt agaagaattc ctgctcaatt acaagctgcc accggcaaat accacaagag
     1681 accagtggta tcatggcact ctgaccagag aagaggcagt agccatacta aaaaagcatg
     1741 ccaaggagct attggccaag caacctgaag tagacacctc cggctgcttt ctggttcgct
     1801 actccgagtc gccggcggcc tcgggattgg ttctaacatt gcttagtgat caggtggtca
     1861 agaattttcg tatttcgcaa gctgatctct accagaatgg taacaaggtg cagtcgggcg
     1921 ggtccaagtt tctgtacatt gatgacggtc cctactggcc aagcgttgaa catctaatag
     1981 cccacttcat gcgtttctcg tacgggttgc cagtgagtct caagtatccg gttccgccac
     2041 agcccaagcc agaggtgcca tcgtttgcca caatacctcg atctaatatg aagccaaaag
     2101 cagcatcgcc cgctaccccg cccactccag tatcgccaca ttcgcatcat caacaccccc
     2161 acgttcccgc cctgaccata acgaagaaaa agcagaagga gaacagtagc tcaatgttca
     2221 acactctgcg actgacgagt ccgaagaaag cgctattcga catgaacagc ctgcgcaaaa
     2281 acaagtccaa gggcaaacgc agtgactccg agagcagtgt gagtggttct ttggccagaa
     2341 cggagcagga gctgcaggca gcggcaccca tgctgaagag cctgtcgttc tcgacggagt
     2401 tctcaacgtt taacgcggat ggcgtaactg gttctggtgc cgccgctgct ggcgaagtgt
     2461 acaacgtgcc caggaacaac acgcccattg agattgacct gccaccgatt gcccaaaaga
     2521 ctgaagccga agttgagtat ttcacgaaaa gtgatgtggc catagagcgc gagcgagcag
     2581 gtcagtggat cggcaacggc taccagccca cagtggacgt gctctccctg ttggatcagc
     2641 agatcaaggc gcccgccgtg gcgcgactaa actcgcttgg tccgaacgca tcaacggaaa
     2701 gtgagatggc cagctatctt catcgcaagt gcagcggtac gccaagcaca ccaagcgcca
     2761 ctgaggtgga ggccgccaag ctgagattct ttattgagcc agaaaaacta gtgctggata
     2821 gagagatcgg gcacggggaa ttcggttcgg tgcacagtgg atggctgctg agaaagagcg
     2881 gtgccggcga ggaatcgcgc ttggaggtgg ccattaagat gcttagcgac gagcacagca
     2941 acaaacagga gttcctgcgc gaagcatcag ttatgatgcg gctggagcac aagtgcattg
     3001 tgcgattgat tggcatttcg aagggtgaaa tgctaatgat ggttcaagag ctagctccgc
     3061 tgggctccat gctgcagtac atactggatc atggtcatga gatcacggcc aatgcggagc
     3121 tgaaagtttg ggcgtcacag attgcgtgtg gcatgcatta tctggaatca cagcattttg
     3181 tgcaccgcga tctggccgcc agaaatatat tgctgaccgc acgccaccag gcgaagatca
     3241 gtgacttcgg catgtcgcga tcactgagac ccggtagcac ggaataccaa ttcacccagg
     3301 gcggacgttg gcccatccgc tggtacgcac cggaaagctt caacctgggc atcttctcgc
     3361 acgccagtga tgtgtggtcc tttggcgtga ccatctggga gatgttttcc ctcggcgcac
     3421 cgccatatgg ggaaatatct aatgtcgatg ccatcaagtt ggtggacagt ggcgaacgcc
     3481 tgccgcagcc caatctctgt ccagcttaca tctatgccgt gatgcagagc tgctggaagg
     3541 aacgacccaa agatcggcca acctttgtct atctcaccga gtttttcgcc cgcgatcctg
     3601 attaccaaaa cctgccggaa ttggtgcaaa cggttcacat tggttctggt tctatgagcg
     3661 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc
     3721 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa
     3781 tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc
     3841 tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt
     3901 ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga
     3961 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag
     4021 gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct
     4081 tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga
     4141 agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga tccaagaaac
     4201 ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca
     4261 aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg
     4321 acctccctag caaactgggg cacaagctta attaagttta aacgcccgct gatcagcctc
     4381 gactgtgcct tctaagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     4441 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4501 gtaaccatta taagctgcaa taaacaagtt aactctagag gatccggtgc gtggtggttt
     4561 catgcttctg ggaacggcaa atgggtttag gattgggaac ccctcatcat ctgttggaat
     4621 atactattca acctacaaaa gtaacgttaa acaacactac tttatatttg atatgaatgg
     4681 ccacaccttt tatgccataa aacatattgt aagagaatac cactcttttt attccttctt
     4741 tccttcttgt acgttttttg ctgtaagtag gtcgtggtgc tggtgttgca gttgaaataa
     4801 cttaaaatat aaatcataaa actcaaacat aaacttgact atttatttat ttattaagaa
     4861 aggaaatata aattataaat tacaacagat cccccgggct gcaggaattc gatagcttgc
     4921 atgcctgcag gtcggccgcc acgaccggtg ccgccaccat cccctgaccc acgcccctga
     4981 cccctcacaa ggagacgacc ttccatgacc gagtacaagc ccacggtgcg cctcgccacc
     5041 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     5101 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     5161 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     5221 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     5281 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     5341 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     5401 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     5461 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5521 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5581 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg
     5641 agcgcacgac cccatggctc cgaccgaagc cacccggggc ggccccgccg accccgcacc
     5701 cgcccccgag gcccaccgac tctagatcaa gcttatcgat accgtcgacc tcgaggccag
     5761 ctagcttgaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5881 atgtatctta tcatgtctgg atcgggtacc caattgcatt cattttatgt ttcaggttca
     5941 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     6001 ttatgatcag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     6061 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6121 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6181 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6241 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6301 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6361 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     8221 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8281 tgatgacggt gaaaacctct g
//
