LOCUS       sequence-376797-        8010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive expression of CRY2PHR-VPR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8010)
  AUTHORS   Huang Z, Wu Y, Allen ME, Pan Y, Kyriakakis P, Lu S, Chang YJ, Wang 
            X, Chien S, Wang Y
  TITLE     Engineering light-controllable CAR T cells for cancer immunotherapy.
  JOURNAL   Sci Adv. 2020 Feb 19;6(8):eaay9209. doi: 10.1126/sciadv.aay9209. 
            eCollection 2020 Feb.
  PUBMED    32128416
REFERENCE   2  (bases 1 to 8010)
  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: "Sci Adv."; 
            date: "2020-02-19"; volume: "6(8):eaay9209. doi"; pages: " 
            10.1126/sciadv.aay9209. eCollection 2020 Feb"
COMMENT     Sequence Label: pCBI-CRY2PHR-VPR
FEATURES             Location/Qualifiers
     source          1..8010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1626..1648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1714..1733
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1774..1821
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3322..3342
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    3388..3412
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3442..3591
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             3616..3636
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4066..4422
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             4441..5010
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     primer_bind     complement(5032..5051)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5097..5152
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5151..5170)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5513..5529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5513..5529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5526..5548)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5537..5553
                     /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(5561..5567,5568..5585,5586..5591))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5606..5627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5686..5881
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5732..5867
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5887..6021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5924..5943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5978..5997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6089..6106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6260..6848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6340..6359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7019..7810,7811..7879))
                     /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     7642..7661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7880..7984)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg gcgcgccrgg gggggggggg gggggggggg
      541 gggggggsgg ggsgrggcgg rgaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggrgggct cgggggrggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gygckgrkcc saaatctgkg nngcggmgcc
     1441 gaamycyggg wggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     1501 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     1561 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     1621 ggcggggttc ggcttctggc gtgtgaccgg cggctctagc tagagcctct gctaaccatg
     1681 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     1741 atcattttgg caaagaattc tgctgccacc atgaaaaggc cggcggccac gaaaaaggcc
     1801 ggccaggcaa aaaagaaaaa gaagatggac aaaaagacga ttgtgtggtt tagacgtgac
     1861 cttcgcattg aggacaaccc tgcgttggca gcggcagcgc atgaaggttc ggtattccct
     1921 gtctttatct ggtgtcccga agaggagggg caattctatc ccggtcgcgc gtcccgatgg
     1981 tggatgaaac agtcgctcgc acatcttagc cagtcgttga aggcgctcgg ctctgatctg
     2041 acgctcatca aaacacacaa tacaatttct gccattctgg attgcattag ggtgactggt
     2101 gccaccaaag ttgtgttcaa tcacctttat gatcccgtaa gcctggtcag ggatcataca
     2161 gtaaaagaga aacttgtaga gaggggaatc agcgtccaat cgtacaatgg ggacttgttg
     2221 tatgaaccgt gggaaatcta ttgtgagaaa ggcaaacctt tcacttcctt caattcgtac
     2281 tggaagaaat gcctcgacat gtccatcgag tcagtcatgc tgccgccacc ttggagactt
     2341 atgcccatta ctgctgcggc agaggcgatc tgggcttgca gcatcgaaga acttggtctg
     2401 gaaaacgaag cggaaaaacc cagtaacgcg ctcctgacgc gagcgtggtc gccaggctgg
     2461 tcgaatgctg ataagctcct taatgagttt atcgagaaac agttgatcga ctatgcgaag
     2521 aatagcaaga aggtggtggg aaattccact tccctcctgt cgccgtatct ccactttggg
     2581 gagattagcg tgaggcatgt gttccaatgt gcgcggatga agcaaatcat ttgggctcgc
     2641 gacaaaaact ctgaaggaga ggagtctgcc gatttgtttt tgagaggaat cggccttagg
     2701 gagtattcac gatacatctg cttcaacttt ccgtttacac atgaacaatc cctcctcagc
     2761 cacctccgct tctttccgtg ggacgcggac gtggacaaat tcaaggcctg gcggcagggg
     2821 cgcacgggat acccgttggt ggatgcgggt atgagagagc tgtgggcaac gggttggatg
     2881 cacaaccgta ttcgagtcat cgttagtagc tttgccgtta agttcctctt gttgccctgg
     2941 aaatggggga tgaaatactt ttgggacacg cttctggatg cggacttgga gtgcgatatt
     3001 cttgggtggc agtacatttc gggttccatc cctgacgggc atgaacttga ccgattggat
     3061 aaccctgcac tgcaaggagc aaagtacgac ccggagggag aatacatcag gcagtggctc
     3121 ccagagctgg cccgtctgcc caccgagtgg attcaccacc cgtgggatgc tcccctgacc
     3181 gtccttaaag cctcaggggt cgaattgggg acgaattacg cgaaacccat cgtcgatatt
     3241 gacaccgcac gagaattgct ggcgaaagcc atctcaagaa cacgcgaagc gcagattatg
     3301 atcggagcag cgccggctag ccccaagaag aagagaaagg tggaggccag ctcgccaggg
     3361 atccgtcgac ttgacgcgtt gatatcaaca agtttgtaca aaaaagcagg ctacaaagag
     3421 gccagcggtt ccggacgggc tgacgcattg gacgattttg atctggatat gctgggaagt
     3481 gacgccctcg atgattttga ccttgacatg cttggttcgg atgcccttga tgactttgac
     3541 ctcgacatgc tcggcagtga cgcccttgat gatttcgacc tggacatgct gattaactct
     3601 agaagttccg gatctccgaa aaagaaacgc aaagttggta gccagtacct gcccgacacc
     3661 gacgaccggc accggatcga ggaaaagcgg aagcggacct acgagacatt caagagcatc
     3721 atgaagaagt cccccttcag cggccccacc gaccctagac ctccacctag aagaatcgcc
     3781 gtgcccagca gatccagcgc cagcgtgcca aaacctgccc cccagcctta ccccttcacc
     3841 agcagcctga gcaccatcaa ctacgacgag ttccctacca tggtgttccc cagcggccag
     3901 atctctcagg cctctgctct ggctccagcc cctcctcagg tgctgcctca ggctcctgct
     3961 cctgcaccag ctccagccat ggtgtctgca ctggctcagg caccagcacc cgtgcctgtg
     4021 ctggctcctg gacctccaca ggctgtggct ccaccagccc ctaaacctac acaggccggc
     4081 gagggcacac tgtctgaagc tctgctgcag ctgcagttcg acgacgagga tctgggagcc
     4141 ctgctgggaa acagcaccga tcctgccgtg ttcaccgacc tggccagcgt ggacaacagc
     4201 gagttccagc agctgctgaa ccagggcatc cctgtggccc ctcacaccac cgagcccatg
     4261 ctgatggaat accccgaggc catcacccgg ctcgtgacag gcgctcagag gcctcctgat
     4321 ccagctcctg cccctctggg agcaccaggc ctgcctaatg gactgctgtc tggcgacgag
     4381 gacttcagct ctatcgccga tatggatttc tcagccttgc tgggctctgg cagcggcagc
     4441 cgggattcca gggaagggat gtttttgccg aagcctgagg ccggctccgc tattagtgac
     4501 gtgtttgagg gccgcgaggt gtgccagcca aaacgaatcc ggccatttca tcctccagga
     4561 agtccatggg ccaaccgccc actccccgcc agcctcgcac caacaccaac cggtccagta
     4621 catgagccag tcgggtcact gaccccggca ccagtccctc agccactgga tccagcgccc
     4681 gcagtgactc ccgaggccag tcacctgttg gaggatcccg atgaagagac gagccaggct
     4741 gtcaaagccc ttcgggagat ggccgatact gtgattcccc agaaggaaga ggctgcaatc
     4801 tgtggccaaa tggacctttc ccatccgccc ccaaggggcc atctggatga gctgacaacc
     4861 acacttgagt ccatgaccga ggatctgaac ctggactcac ccctgacccc ggaattgaac
     4921 gagattctgg ataccttcct gaacgacgag tgcctcttgc atgccatgca tatcagcaca
     4981 ggactgtcca tcttcgacac atctctgttt taaagtgcat gagtcgacgg atctttttcc
     5041 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     5101 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     5161 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     5221 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     5281 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     5341 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     5401 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     5461 tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     5521 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5581 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5641 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     5701 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5761 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5821 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5881 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5941 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6001 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6061 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6661 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7141 gtgctgcaat gataccgcgt gacccacgct caccggctcc agatttatca gcaataaacc
     7201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7321 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7621 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7981 cgcgcacatt tccccgaaaa gtgccacctg
//
