LOCUS       sequence-479873-        8198 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  Retroviral expression construct encoding CRY2clust fused to two 
            copies of the C-terminal tail of human STING.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8198)
  AUTHORS   Dou Y, Chen R, Liu S, Lee YT, Jing J, Liu X, Ke Y, Wang R, Zhou Y, 
            Huang Y
  TITLE     Optogenetic engineering of STING signaling allows remote 
            immunomodulation to enhance cancer immunotherapy.
  JOURNAL   Nat Commun. 2023 Sep 6;14(1):5461. doi: 10.1038/s41467-023-41164-2.
  PUBMED    37673917
REFERENCE   2  (bases 1 to 8198)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-09-6"; volume: "14(1):5461. doi"; pages: " 
            10.1038/s41467-023-41164-2"
COMMENT     Sequence Label: pMSCV-GFP-IRES-CRY2clust-pLxIS
FEATURES             Location/Qualifiers
     source          1..8198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(123..227)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     295..313
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(351..373)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     473..492
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     686..708
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     836..855
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             1055..1571
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1635..1976
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2043..2459
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2389..2411
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2427..2443
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4286..4852
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4302..4854
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4446..4463)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4673..4692
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4866..4868,4869..4871,4872..5585)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4866..4868,4869..4871,4872..5582)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4911..4932)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5172..5191)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5519..5540
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5647..6161
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6323..6339
                     /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(6347..6353,6354..6371,6372..6377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6392..6413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6530..6547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6701..7289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6781..6800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7460..8198,1..53,54..122))
                     /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     8083..8102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
       61 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
      121 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
      181 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
      241 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
      301 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
      361 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
      421 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
      481 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
      541 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      601 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      661 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga
      721 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
      781 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
      841 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
      901 gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
      961 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
     1021 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     1081 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     1141 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     1201 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     1261 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     1321 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     1381 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     1441 atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga
     1501 atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg
     1561 ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc
     1621 ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg
     1681 tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta
     1741 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     1801 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     1861 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     1921 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     1981 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     2041 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     2101 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     2161 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     2221 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     2281 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     2341 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
     2401 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
     2461 gaattagatc catgaagatg gacaaaaaga ccatcgtctg gtttcggaga gatttgagaa
     2521 tagaagataa tcccgcgctc gccgccgcgg cccacgaggg ttccgtcttc cccgttttca
     2581 tttggtgtcc tgaagaagaa ggccagtttt atcccggaag ggcctctagg tggtggatga
     2641 agcaaagtct ggcccatctt agccagtcac tgaaagcact gggcagtgat cttaccctga
     2701 tcaagacaca caataccatc tctgccatcc tcgactgcat cagggtgacc ggcgcaacga
     2761 aagtcgtgtt taaccacctg tacgatccag ttagtctggt gcgcgaccac actgtgaagg
     2821 agaagctggt ggaacggggg atcagtgtgc agagctacaa cggggacctt ctgtacgagc
     2881 catgggagat ctattgcgag aaagggaaac cgttcacctc cttcaacagt tactggaaga
     2941 aatgtttgga tatgtcaata gagtccgtta tgttgccccc tccctggaga ctgatgccga
     3001 ttactgctgc tgcagaggcc atctgggcct gctccatcga ggaactcggt ctggaaaatg
     3061 aagcagaaaa gccaagcaat gcacttctca ctagagcctg gagccccggc tggtctaatg
     3121 ccgacaagct gcttaacgag ttcatcgaaa aacaactgat tgactacgcg aagaactcca
     3181 agaaagtggt aggtaactca actagcttgc tctctccata tctccatttt ggcgagattt
     3241 ctgtccgcca tgtatttcag tgcgctcgga tgaaacagat tatctgggct cgcgataaaa
     3301 acagcgaagg cgaagaaagc gccgatctgt tcctgcgagg gatcggactt cgggaatact
     3361 cccggtatat atgtttcaac tttccattca cacacgagca gagtctgttg tcccacctca
     3421 ggttcttccc ctgggacgcc gatgtcgaca aattcaaggc atggagacag ggaaggacag
     3481 ggtacccact cgtggatgct ggcatgagag agctctgggc tacaggctgg atgcacaacc
     3541 gcatccgggt aatcgtgtcc tcatttgctg tcaagtttct gctcctgcct tggaaatggg
     3601 gaatgaagta cttttgggat acccttctcg acgccgactt ggagtgtgac attctgggat
     3661 ggcaatatat tagcgggtca attcctgacg gccatgagtt ggacaggttg gacaatccgg
     3721 ccttgcaggg agctaagtat gatcccgaag gagagtatat tcgacagtgg ctccccgagc
     3781 tggcccgact tcctacggag tggattcacc atccttggga cgcaccactg acagtgctca
     3841 aggcaagcgg ggtggagctg ggcaccaatt acgctaagcc tatagttgat atagatacag
     3901 cacgcgagct gctggctaaa gcgatctctc gcactcggga ggcgcagatt atgatcggtg
     3961 ctgccgcaag ggaccctcct gatcttgaca atagtggggg agttactatg aacgcaccta
     4021 tgacatctgt ggcccctccc ccttccgttc ttagtcagga accccgcctt ttgataagtg
     4081 gcatggacca accactccct ctgaggacag accttattgt gactatgaat gcccccatga
     4141 cctctgtagc cccaccacca agcgtactta gtcaagagcc aagactcctg atatctggta
     4201 tggaccaacc actgccactc agaactgatc tgatttaagc ggccgcgttt aaacgtcgac
     4261 ggtatcgata agcttgatat cgaattccgc ccccccccct aacgttactg gccgaagccg
     4321 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     4381 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     4441 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     4501 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     4561 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     4621 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     4681 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     4741 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     4801 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     4861 caaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4921 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     4981 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     5041 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     5101 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     5161 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     5221 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5281 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     5341 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     5401 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5461 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     5521 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5581 agtaatgaat taattaagaa ttcgataaaa taaaagattt tatttagtct ccagaaaaag
     5641 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     5701 aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga
     5761 gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
     5821 gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca
     5881 gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa
     5941 tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc
     6001 acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta
     6061 tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc
     6121 tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag
     6181 ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc
     6241 actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa
     6301 agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     6361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     6421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     6481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     6541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7561 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     7801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8161 tcttaccgct gttgagatcc agttcgatgt aacccact
//
