LOCUS       sequence-369337-        6957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual secreted luciferase reporter (EF1alpha-Gaussia luciferase, 
            CMV-Cypridina luciferase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6957)
  AUTHORS   Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung 
            J, Zhang F
  TITLE     RNA editing with CRISPR-Cas13.
  JOURNAL   Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
  PUBMED    29070703
REFERENCE   2  (bases 1 to 6957)
  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"; journalName: "Science. 
            2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180."
COMMENT     Sequence Label: pC0037
FEATURES             Location/Qualifiers
     source          1..6957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(136..724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(216..235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(895..1686,1687..1755))
                     /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     1518..1537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1756..1860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1928..1946
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1984..2006)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2106..2125
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2319..2341
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2333..2350
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2334..2350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2373..2421
                     /note="synthetic polyadenylation signal"
     CDS             complement(2422..2979)
                     /codon_start=1
                     /product="secreted Gaussia luciferase"
                     /label=hGLuc
                     /note="human codon-optimized"
                     /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG
                     KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA
                     QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ
                     RCATFASKIQGQVDKIKGAGGD"
     regulatory      2976..2985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3004..4182)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3013..3951
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3036..3056)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     enhancer        4295..4674
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4675..4878
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4829..4849
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4923..4942
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4923..4941
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4967..6628
                     /codon_start=1
                     /product="secreted Cypridina luciferase"
                     /label=CLuc
                     /note="contains 30 codon substitutions for efficient 
                     translation in mammalian cells"
                     /translation="MKTLILAVALVYCATVHCQDCPYEPDPPNTVPTSCEAKEGECIDS
                     SCGTCTRDILSDGLCENKPGKTCCRMCQYVIECRVEAAGWFRTFYGKRFQFQEPGTYVL
                     GQGTKGGDWKVSITLENLDGTKGAVLTKTRLEVAGDIIDIAQATENPITVNGGADPIIA
                     NPYTIGEVTIAVVEMPGFNITVIEFFKLIVIDILGGRSVRIAPDTANKGMISGLCGDLK
                     MMEDTDFTSDPEQLAIQPKINQEFDGCPLYGNPDDVAYCKGLLEPYKDSCRNPINFYYY
                     TISCAFARCMGGDERASHVLLDYRETCAAPETRGTCVLSGHTFYDTFDKARYQFQGPCK
                     EILMAADCFWNTWDVKVSHRNVDSYTEVEKVRIRKQSTVVELIVDGKQILVGGEAVSVP
                     YSSQNTSIYWQDGDILTTAILPEALVVKFNFKQLLVVHIRDPFDGKTCGICGNYNQDFS
                     DDSFDAEGACDLTPNPPGCTEEQKPEAERLCNSLFAGQSDLDQKCNVCHKPDRVERCMY
                     EYCLRGQQGFCDHAWEFKKECYIKHGDTLEVPDECK"
     polyA_signal    6629..6677
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6691..6707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6691..6707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6704..6726)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6715..6731
                     /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(6739..6745,6746..6763,6764..6769))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6784..6805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6922..6939)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
       61 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      121 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      181 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      241 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      301 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      361 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      421 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      481 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      541 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      601 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      661 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      841 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      961 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1021 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1081 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1261 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1321 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1381 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1441 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1501 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1561 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1621 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1681 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1741 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1801 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1861 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1921 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     1981 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2041 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2101 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     2161 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2221 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2281 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2341 gacggccagt gaattcgagc tcggtacccg ggcacacaaa aaaccaacac acagatgtaa
     2401 tgaaaataaa gatattttat tttagtcacc accggccccc ttgatcttgt ccacctggcc
     2461 ctggatcttg ctggcaaagg tcgcacagcg ttgcggcagc cacttcttga gcaggtcaga
     2521 acactgcacg ttggcaagcc ctttgaggca gccagttgtg cagtccacac acagatcgac
     2581 ctgtgcgatg aactgctcca tgggctccaa gtccttgaac ccaggaatct caggaatgtc
     2641 gacgatcgcc tcgcctatgc cgccctgtgc ggactctttg tcgccttcgt aggtgtggca
     2701 gcgtcctggg atgaacttct tcatcttggg cgtgcacttg atgtgggaca ggcagatcag
     2761 acagcccctg gtgcagccag ctttccgggc attggcttcc atctctttga gcacctccag
     2821 cggcagcttc ttgccgggca acttcccgcg gtcagcatcg agatccgtgg tcgcgaagtt
     2881 gctggccacg gccacgatgt tgaagtcttc gttgttctcg gtgggcttgg cctcggccac
     2941 agcgatgcag atcagggcaa acagaacttt gactcccatg gtggctctag aggatcccgt
     3001 acgtcacgac acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat
     3061 gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc
     3121 tggcctaact tcagtctcca cccactcagt gtggggaaac tccatcgcat aaaacccctc
     3181 cccccaacct aaagacgacg tactccaaaa gctcgagaac taatcgaggt gcctggacgg
     3241 cgcccggtac tccgtggagt cacatgaagc gacggctgag gacggaaagg cccttttcct
     3301 ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc gtcctccatt ttgagctccc
     3361 tgcagcaggg ccgggaagcg gccatctttc cgctcacgca actggtgccg accgggccag
     3421 ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg ccaggcacca gagcaggccg
     3481 gccagcttga gactaccccc gtccgattct cggtggccgc gctcgcaggc cccgcctcgc
     3541 cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc gcggccccaa aaaccgaaat
     3601 accagtgtgc agatcttggc ccgcatttac aagactatct tgccagaaaa aaagcgtcgc
     3661 agcaggtcat caaaaatttt aaatggctag agacttatcg aaagcagcga gacaggcgcg
     3721 aaggtgccac cagattcgca cgcggcggcc ccagcgccca ggccaggcct caactcaagc
     3781 acgaggcgaa ggggctcctt aagcgcaagg cctcgaactc tcccacccac ttccaacccg
     3841 aagctcggga tcaagaatca cgtactgcag ccaggtggaa gtaattcaag gcacgcaagg
     3901 gccataaccc gtaaagaggc caggcccgcg ggaaccacac acggcactta cctgtgttct
     3961 ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc
     4021 tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc
     4081 gcgccacctt ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac
     4141 tgtgggcgat gtgcgctctg cccactgacg ggcaccggag ccaattccca ctcctttcaa
     4201 gacctagaag gtccattagc tgcaaagatt cctctctgtt taaaacttta tccatcttgc
     4261 atcagcttcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     4321 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     4381 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     4441 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     4501 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     4561 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     4621 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     4681 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     4741 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     4801 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     4861 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     4921 attaatacga ctcactatag ggagacccaa gcttggtacc gccaccatga agaccttaat
     4981 tcttgccgtt gcattagtct actgcgccac tgttcattgc caggactgtc cttacgaacc
     5041 tgatccacca aacacagttc caacttcctg tgaagctaaa gaaggagaat gtattgatag
     5101 cagctgtggc acctgcacga gagacatact atcagatgga ctgtgtgaaa ataaaccagg
     5161 aaaaacatgt tgtcgaatgt gtcagtatgt aattgaatgc agagtagagg ccgcaggatg
     5221 gtttagaaca ttctatggaa agagattcca gttccaggaa cctggtacat acgtgttggg
     5281 tcaaggaacc aagggcggcg actggaaggt gtccatcacc ctggagaacc tggatggaac
     5341 caagggggct gtgctgacca agacaagact ggaagtggct ggagacatca ttgacatcgc
     5401 tcaagctact gagaatccca tcactgtaaa cggtggagct gaccctatca tcgccaaccc
     5461 gtacaccatc ggcgaggtca ccatcgctgt tgttgagatg ccaggcttca acatcaccgt
     5521 cattgagttc ttcaaactga tcgtgatcga catcctcgga ggaagatctg taagaatcgc
     5581 cccagacaca gcaaacaaag gaatgatctc tggcctctgt ggagatctta aaatgatgga
     5641 agatacagac ttcacttcag atccagaaca actcgctatt cagcctaaga tcaaccagga
     5701 gtttgacggt tgtccactct atggaaatcc tgatgacgtt gcatactgca aaggtcttct
     5761 ggagccgtac aaggacagct gccgcaaccc catcaacttc tactactaca ccatctcctg
     5821 cgccttcgcc cgctgtatgg gtggagacga gcgagcctca cacgtgctgc ttgactacag
     5881 ggagacgtgc gctgctcccg aaactagagg aacctgcgtt ttgtctggac atactttcta
     5941 cgatacattt gacaaagcaa gataccaatt ccagggtccc tgcaaggaga ttcttatggc
     6001 cgccgactgt ttctggaaca cttgggatgt gaaggtttca cacaggaatg ttgactctta
     6061 cactgaagta gagaaagtac gaatcaggaa acaatcgact gtagtagaac tcattgttga
     6121 tggaaaacag attctggttg gaggagaagc cgtgtccgtc ccgtacagct ctcagaacac
     6181 ttccatctac tggcaagatg gtgacatact gactacagcc atcctacctg aagctctggt
     6241 ggtcaagttc aacttcaagc aactgctcgt cgtacatatt agagatccat tcgatggtaa
     6301 gacttgcggt atttgcggta actacaacca ggatttcagt gatgattctt ttgatgctga
     6361 aggagcctgt gatctgaccc ccaacccacc gggatgcacc gaagaacaga aacctgaagc
     6421 tgaacgactc tgcaatagtc tcttcgccgg tcaaagtgat cttgatcaga aatgtaacgt
     6481 gtgccacaag cctgaccgtg tcgaacgatg catgtacgag tattgcctga ggggacaaca
     6541 gggtttctgt gaccacgcat gggagttcaa gaaagaatgc tacataaagc atggagacac
     6601 cctagaagta ccagatgaat gcaaataaaa taaaatatct ttattttcat tacatctgtg
     6661 tgttggtttt ttgtgtgtgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggct
//
