LOCUS       Exported                7855 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Inducible expression of Stat92E-GFP in S2 cells.
ACCESSION   .
VERSION     .
KEYWORDS    pMT Stat92E-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7855)
  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 7855)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7855
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     130..152
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     144..161
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     145..161
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        167..593
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             628..645
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2998..3708
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3025..3053)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3648..3668
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3731..3748)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3750..3884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             4359..4958
                     /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(4359..4378)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4855..4875
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5091..5107)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5092..5108)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5123..5257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5400..5416)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5400..5416)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5413..5435)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5424..5440
                     /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(5448..5478)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5493..5514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5631..5648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5802..6390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5882..5901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6561..7421)
                     /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     7184..7203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7422..7526)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7594..7612
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7650..7672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7772..7791
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
       61 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      121 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattaatt cgttgcagga
      181 caggatgtgg tgcccgatgt gactagctct ttgctgcagg ccgtcctatc ctctggttcc
      241 gataagagac ccagaactcc ggccccccac cgcccaccgc cacccccata catatgtggt
      301 acgcaagtaa gagtgcctgc gcatgcccca tgtgccccac caagagcttt gcatcccata
      361 caagtcccca aagtggagaa ccgaaccaat tcttcgcggg cagaacaaaa gcttctgcac
      421 acgtctccac tcgaatttgg agccggccgg cgtgtgcaaa agaggtgaat cgaacgaaag
      481 acccgtgtgt aaagccgcgt ttccaaaatg tataaaaccg agagcatctg gccaatgtgc
      541 atcagttgtg gtcagcagca aaatcaagtg aatcatctca gtgcaactaa aggggggatc
      601 tagatcgggg tacctactag tgaaatgcac catcaccatc atcacgaaga tcaagttgac
      661 ccaagactta ttgacggaaa aggaggagga ggccggccca tgagcttgtg gaagcgcatc
      721 tccagccatg tcgactgcga gcagcgtatg gcggcttact acgaggagaa gggtatgctc
      781 gagctgcgcc tctgcttggc accctggatc gaagacagga taatgtccga acaaataacg
      841 cccaacacta ccgatcagtt agagcgcgtg gccctaaagt tcaacgaaga tctgcagcaa
      901 aagcttctat ccacgcgcac cgccagcgac caggccctca agttccgggt ggtggagctt
      961 tgcgccctca tccaacgcat ctccgccgtt gagctgtaca cgcatctgcg cagcggtcta
     1021 caaaaagagt tgcagttggt caccgagaag agcgtggctg caaccgcagg ccaatcaatg
     1081 ccgctaaatc cctacaacat gaacaacacg cccatggtta ccgggtacat ggtggacccc
     1141 agtgatcttc tggcggtgtc caacagctgc aatcctccag ttgttcaggg cataggacct
     1201 atccacaatg tgcaaaatac aggcattgcc tccccagccc tcggtatggt cacacccaag
     1261 gtggagctgt acgaggtgca acatcagatc atgcagtccc tcaatgagtt tggcaactgt
     1321 gccaatgcct tgaagctgct tgcccaaaac tacagttaca tgctaaactc cacatcctcg
     1381 ccgaacgcag aagctgccta cagaagtctt atcgatgaaa aggcggccat cgtactcaca
     1441 atgcgtcgca gctttatgta ctacgaatcg ctgcacgaga tggtcataca cgagctcaag
     1501 aactggcgcc accaacaagc gcaagccgga aacggagctc cctttaatga gggctccctg
     1561 gatgatatcc agcgctgctt tgagatgctc gagagcttta tcgcacacat gctggctgcc
     1621 gttaaggagc tgatgcgcgt acgtctggtc accgaagagc cggagctcac acatctgctt
     1681 gaacaggtgc agaacgcgca gaagaacctg gtatgctctg cctttatcgt cgacaaacag
     1741 cccccgcaag tgatgaagac caacacacgc ttcgcagcgt ctgtgcgttg gctaatcggt
     1801 tctcagctgg gcattcacaa caatccaccc acagtcgagt gcatcataat gtcagagatc
     1861 cagtcgcaac ggttcgttac gcgcaataca cagatggata atagtagcct atccggtcaa
     1921 tcatcgggcg agattcaaaa cgcctctagc actatggagt atcagcaaaa caaccatgtt
     1981 ttctccgcca gcttccgaaa catgcagctg aagaagatca agcgggcaga aaagaagggc
     2041 actgagagcg tgatggacga gaagttcgcc ctgttcttct acaccaccac caccgtgaac
     2101 gatttccaga tccgtgtgtg gaccctgtcc ttaccggtgg tggttattgt gcacggcaac
     2161 caggagcccc agtcttgggc taccattact tgggacaatg catttgcgga gattgttcgc
     2221 gatcccttca tgattaccga ccgcgttacc tgggcccagc tgtcagtcgc gttgaacatc
     2281 aaattcggat cgtgcacagg gcgttctctg accattgata accttgattt cctatacgag
     2341 aaactccagc gtgaggagcg gtctgagtac attacgtgga accaattctg taaggagccc
     2401 atgcccgatc ggtcctttac attctgggag tggttctttg ccatcatgaa actcaccaaa
     2461 gatcacatgt tgggcatgtg gaaagcaggc tgcattatgg gcttcatcaa caagaccaag
     2521 gctcagactg atctgctgcg ttcagtctat ggtatcggca ctttcctgct ccgtttctcc
     2581 gacagcgagc tcggtggagt cactatcgcc tacgtaaacg aaaatggact ggtcaccatg
     2641 ctagcgccat ggactgcacg ggatttccag gtgctgaacc tggccgatcg cattcgagat
     2701 ctggacgtgc tttgctggct gcaccctagc gaccgcaatg cgagtcccgt gaagagggac
     2761 gtcgccttcg gtgagttcta ctcaaagcgt caagatcctg tgaccggtta tgtgaagagc
     2821 acattgcatg ttcacgtttg tagaaacgga gagaatggat ctactagtgg aacaccgcat
     2881 catgctcagg aaagcatgca actgggaaat ggtgactttg gaatggcgga cttcgatacg
     2941 attacaaact ttgagaactt tgggcgcgcc ggcggccgct cgagtctaga gggccccagc
     3001 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     3061 aatgggcaca aattttctgt ccgtggagag ggtgaaggtg atgctacaaa cggaaaactc
     3121 acccttaaat ttatttgcac tactggaaaa ctacctgttc cgtggccaac acttgtcact
     3181 actctgacct atggtgttca atgcttttcc cgttatccgg atcacatgaa acggcatgac
     3241 tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc tttcaaagat
     3301 gacgggacct acaagacgcg tgctgaagtc aagtttgaag gtgataccct tgttaatcgt
     3361 atcgagttaa agggtattga ttttaaagaa gatggaaaca ttcttggaca caaactcgag
     3421 tacaacttta actcacacaa tgtatacatc acggcagaca aacaaaagaa tggaatcaaa
     3481 gctaacttca aaattcgcca caacgttgaa gatggttccg ttcaactagc agaccattat
     3541 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcg
     3601 acacaatctg tcctttcgaa agatcccaac gaaaagcgtg accacatggt ccttcttgag
     3661 tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaata ggtttaaacc
     3721 cgctgatcag cctcgactgt gccttctaag atccagacat gataagatac attgatgagt
     3781 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3841 ctattgcttt atttgtaacc attataagct gcaataaaca agttaactct agaggatccg
     3901 gtgcgtggtg gtttcatgct tctgggaacg gcaaatgggt ttaggattgg gaacccctca
     3961 tcatctgttg gaatatacta ttcaacctac aaaagtaacg ttaaacaaca ctactttata
     4021 tttgatatga atggccacac cttttatgcc ataaaacata ttgtaagaga ataccactct
     4081 ttttattcct tctttccttc ttgtacgttt tttgctgtaa gtaggtcgtg gtgctggtgt
     4141 tgcagttgaa ataacttaaa atataaatca taaaactcaa acataaactt gactatttat
     4201 ttatttatta agaaaggaaa tataaattat aaattacaac agatcccccg ggctgcagga
     4261 attcgatagc ttgcatgcct gcaggtcggc cgccacgacc ggtgccgcca ccatcccctg
     4321 acccacgccc ctgacccctc acaaggagac gaccttccat gaccgagtac aagcccacgg
     4381 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     4441 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     4501 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4561 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4621 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4681 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4741 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4801 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     4861 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     4921 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc
     4981 gcccgaccga aaggagcgca cgaccccatg gctccgaccg aagccacccg gggcggcccc
     5041 gccgaccccg cacccgcccc cgaggcccac cgactctaga tcaagcttat cgataccgtc
     5101 gacctcgagg ccagctagct tgaacttgtt tattgcagct tataatggtt acaaataaag
     5161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5221 gtccaaactc atcaatgtat cttatcatgt ctggatcggg tacccaattg cattcatttt
     5281 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     5341 tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     5401 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5461 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5521 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5581 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5641 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5701 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5761 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5821 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5881 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5941 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6001 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6061 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6121 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6181 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6241 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6301 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6361 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6421 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6481 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6541 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6601 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6661 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6721 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6781 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6841 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6901 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6961 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7021 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7081 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7141 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7201 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7261 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7321 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7381 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7441 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7501 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7561 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7621 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     7681 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     7741 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     7801 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catca
//
