LOCUS       sequence-224817-        7670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid for measuring protease activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7670)
  AUTHORS   Gao XJ, Chong LS, Kim MS, Elowitz MB
  TITLE     Programmable protein circuits in living cells.
  JOURNAL   Science. 2018 Sep 21;361(6408):1252-1258. doi: 
            10.1126/science.aat5062.
  PUBMED    30237357
REFERENCE   2  (bases 1 to 7670)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2018-09-21- 21"; volume: "361"; 
            issue: "6408"; pages: "1252-1258"
COMMENT     Sequence Label: PB-CMV-TO-Casp3te
FEATURES             Location/Qualifiers
     source          1..7670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1409..1788
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1789..1992
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1943..1963
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1994..2012
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2015..2033
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2043..2067
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2159..2168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2693..2713
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(3050..3067)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3056..3263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3296..3316)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3301..3630
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3481..3616
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3543..3562
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3678..3725
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3744..4118
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(4178..4198)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    4240..4361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4277..4296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4331..4350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5124..5158
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5649..5665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5649..5665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5662..5684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5673..5689
                     /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(5697..5703,5704..5721,5722..5727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5742..5763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5880..5897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6051..6639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6131..6150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6810..7601,7602..7670))
                     /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     7433..7452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtaggcgttt tgcgctgctt
     1381 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     1441 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     1501 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1561 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1621 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1681 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1741 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1801 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1861 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1921 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1981 ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga
     2041 cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     2101 catagaagac accgggaccg atccagcctc cggactctag cgtttaaact taagcttcgc
     2161 caccatggga tccgaaaaca ctgagaacag cgtcgatagc aagtccatta agaacctcga
     2221 acctaaaatc atacatggca gtgagagcat ggatagcggc ataagccttg acaactccta
     2281 taaaatggac tacccggaga tgggtctgtg cataataatc aacaacaaga acttccacaa
     2341 atccacgggg atgacaagta ggtctggaac cgatgtcgat gcggcaaact tgcgcgagac
     2401 ctttagaaac ctcaagtacg aggtacgcaa taaaaacgat ctcactcgcg aggaaatagt
     2461 agagctgatg cgagatgtca gtaaggaaga tcattccaag agatcctcct tcgtctgcgt
     2521 cttgttgagt cacggagagg aagggatcat attcggtacg aacggcccag tggatttgaa
     2581 aaagataacc aatttcttta gaggtgaccg gtgccgctct ctcacaggaa agccaaaact
     2641 ctttattatc caagcatgta ggggtacgga actggattgt ggaatcgaaa ctgagaactt
     2701 gtacttccag tccggggtag atgacgacat ggcctgccat aaaatccctg tcgaggccga
     2761 ttttctctac gcctactcta ctgcgcctgg gtattacagc tggagaaaca gcaaggatgg
     2821 ttcctggttt atacaaagcc tgtgtgctat gctcaaacag tatgcggata agctggagtt
     2881 catgcatatc cttaccagag taaatcgcaa agtggccaca gagtttgagt cattttcttt
     2941 tgacgcaaca tttcacgcca agaagcagat tccttgtata gtatctatgc tgacgaaaga
     3001 gctgtacttc tatcacagct agcagctagc gaattcgaat ttaaatcggc ctcgactgtg
     3061 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3121 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3181 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3241 gacaatagca ggcatgctgg ggacgcggcc gcaatttaac gcgaattaat tctgtggaat
     3301 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3361 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3421 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3481 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3541 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3601 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3661 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     3721 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     3781 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
     3841 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
     3901 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
     3961 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
     4021 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
     4081 gcaactgcgt gcacttcgtg gccgaggagc aggactgagt cgacaatcaa cctctggatt
     4141 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     4201 gatacgctgc tttaatgcct ttgtatcatg cgttaactaa acttgtttat tgcagcttat
     4261 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4321 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gaattgactc
     4381 aaatgatgtc aattagtcta tcagaagctc atctggtctc ccttccgggg gacaagacat
     4441 ccctgtttaa tatttaaaca gcagtgttcc caaactgggt tcttatatcc cttgctctgg
     4501 tcaaccaggt tgcagggttt cctgtcctca caggaacgaa gtccctaaag aaacagtggc
     4561 agccaggttt agccccggaa ttgactggat tcctttttta gggcccattg gtatggcttt
     4621 ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa
     4681 agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga
     4741 tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc cccctagcgg
     4801 gggagggacg taattacatc cctgggggct ttgggggggg gctgtccctg atatctataa
     4861 caagaaaata tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt
     4921 atcgtatgag ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac
     4981 gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga
     5041 gctccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa
     5101 gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt
     5161 aatctagctg catcaggatc atatcgtcgg gtcttttttc cggctcagtc atcgcccaag
     5221 ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg aggttgaagc
     5281 ggcatggaaa gagtttgccg aggatgactg ctgctgcatt gacgttgagc gaaaacgcac
     5341 gtttaccatg atgattcggg aaggtgtggc catgcacgcc tttaacggtg aactgttcgt
     5401 tcaggccacc tgggatacca gttcgtcgcg gcttttccgg acacagttcc ggatggtcag
     5461 cccgaagcgc atcagcaacc cgaacaatac cggcgacagc cggaactgcc gtgccggtgt
     5521 gcagattaat gacagcggtg cggcgctggg atattacgtc agcgaggacg ggtatcctgg
     5581 ctggatgccg cagaaatgga catggatacc ccgtgagtta cccggcgggc gcgcttggcg
     5641 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5701 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5761 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5821 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5881 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5941 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6001 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6061 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6541 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6601 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6661 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6721 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6781 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6841 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6901 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6961 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7021 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7081 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7141 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7201 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7261 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7321 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7381 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7441 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7501 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7561 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7621 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
//
