LOCUS       sequence-176067-        7885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A lenti-viral vector expressing photoswitchable CDK5 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7885)
  AUTHORS   Zhou XX, Fan LZ, Li P, Shen K, Lin MZ
  TITLE     Optical control of cell signaling by single-chain photoswitchable 
            kinases.
  JOURNAL   Science. 2017 Feb 24;355(6327):836-842. doi: 
            10.1126/science.aah3605.
  PUBMED    28232577
REFERENCE   2  (bases 1 to 7885)
  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: "2017-02-24- 24"; volume: "355"; 
            issue: "6327"; pages: "836-842"
COMMENT     Sequence Label: pLL3.7m-psCDK5
FEATURES             Location/Qualifiers
     source          1..7885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        315..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             531..711
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    758..883
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1380..1613
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1798..1842
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2140..2257
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2316..2628
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2547..2567
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2644..2660
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2645..2661
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2703..2736)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        2794..3097
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3098..3301
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3298..3322
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3427..3443
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3427..3443
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3454..3472
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3454..3471
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3495..3521
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4830..5498
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="SVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSIDLKVKE
                     GGPLPFAYDILTTAFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNATN
                     DITLDGDCYIYEIRFRGTNFPANGPVMQKRTVKWEPSTENLYVRDGVLKGDVVMALSLE
                     GGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQA
                     K"
     LTR             5838..6018
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6080..6668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6160..6179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6839..7630,7631..7699))
                     /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     7462..7481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7700..7804)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
       61 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      121 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      181 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      241 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      301 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      361 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      421 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      481 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
      541 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      601 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      661 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
      721 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
      781 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      841 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      901 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      961 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     1021 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     1081 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     1141 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     1201 taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg
     1261 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1321 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1381 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1441 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1501 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1561 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1621 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1681 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1741 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1801 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1861 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1921 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1981 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2041 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2101 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2161 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2221 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2281 agatccagtt tggttagtac cgggcccgct ctagagatcc gacgcgccat ctctaggccc
     2341 gcgccggccc cctcgcacag acttgtggga gaagctcggc tactcccctg ccccggttaa
     2401 tttgcatata atatttccta gtaactatag aggcttaatg tgcgataaaa gacagataat
     2461 ctgttctttt taatactagc tacattttac atgataggct tggatttcta taagagatac
     2521 aaatactaaa ttattatttt aaaaaacagc acaaaaggaa actcacccta actgtaaagt
     2581 aattgtgtgt tttgagacta taaatatccc ttggagaaaa gccttgttaa cgcgcggtga
     2641 ccctcgaggt cgacggtatc gataagctcg cttcacgaga ttccagcagg tcgagggacc
     2701 taataacttc gtatagcata cattatacga agttatatta agggttccaa gcttaagcgg
     2761 ccgcgtggat aaccgtatta ccgccatgca tgtcgttaca taacttacgg taaatggccc
     2821 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2881 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2941 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     3001 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     3061 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     3121 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     3181 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     3241 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     3301 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     3361 aagacaccgg gaccgatcca gcctccggac tctagcctag gccgcgggac ggataacaat
     3421 ttcacacagg aaacagctat gaccattagg cctatttagg tgacactata gaacaagttt
     3481 gtacagccac catgtatccc tacgatgtgc ccgattacgc tagcgtgatc aagcccgaca
     3541 tgaaaatcaa gctgaggatg gagggtgccg tgaacggtca cccgtttgcg atcgagggcg
     3601 tgggactggg caagccgttc gagggaaagc aaagcatcga cctcaaggtg aaggagggcg
     3661 gaccgctgcc cttcgcctac gacattctga ccactgcctt ctgctacggt aatcgcgtct
     3721 tcgcaaagta tcccgaaaac atcgtggatt actttaagca gagcttcccg gagggttact
     3781 cctgggagcg atccatgaac tacgaggacg gagggatctg caacgcgacg aacgatataa
     3841 ccttggacgg cgactgctac atttacgaaa ttcggttccg cggcaccaat ttcccagcga
     3901 atggccctgt aatgcaaaaa cgcacggtaa agtgggagcc cagcaccgag aacctgtacg
     3961 tccgcgacgg agtcctgaag ggagacgtgg tcatggcgct ttcattggaa ggtggcgggc
     4021 actacagatg cgatttcaag acgacctata aggcgaaaaa ggtggtccaa ctgccggact
     4081 accatttcgt cgaccaccac atcgagatca agagccacga caaagactat agcaacgtga
     4141 atctccacga gcatgccgag gcccacagcg agctgcccag gcaggccaag ggccttaagg
     4201 gatccggaca gaaatacgag aaactggaaa agattgggga aggcacctac ggaactgtgt
     4261 tcaaggccaa aaaccgggag actcatgaga tcgtggctct gaaacgggtg aggctggatg
     4321 acgatgatga gggtgtgccg agttccgccc tccgggagat ctgcctactc aaggagctga
     4381 agcacaagaa catcgtcagg cttcatgacg tcctgcacag cgacaagaag ctgactttgg
     4441 tttttgaatt ctgtgaccag gacctgaaga agtattttga cagttgcaat ggtgacctcg
     4501 atcctgagat tgtaaagtca ttcctcttcc agctactaaa agggctggga ttctgtcata
     4561 gccgcaatgt gctacacagg gacctgaagc cccagaacct gctaataaac aggaatgggg
     4621 agctgaaatt ggctgatttt ggcctggctc gagcctttgg gattcccgtc cgctgttact
     4681 cagctgaggt ggtcacactg tggtaccgcc caccggatgt cctctttggg gccaagctgt
     4741 actccacgtc catcgacatg tggtcagccg gctgcatctt tgcagagctg gccaatgctg
     4801 ggcggcctct ttttcccggc accggtggca gtgtgattaa accagacatg aagatcaagc
     4861 tgcgtatgga aggcgctgta aatggacacc cgttcgcgat tgaaggagtt ggccttggga
     4921 agcctttcga gggaaaacag agtattgacc ttaaagtcaa agaaggcgga cctctgcctt
     4981 tcgcctatga catcttgaca actgctttct gttacggcaa cagggtattc gccaaatacc
     5041 cagaaaatat agtagactat ttcaagcagt cgtttcctga gggctactct tgggaacgaa
     5101 gcatgaatta cgaagacggg ggcatttgta acgcgacaaa cgacataacc ctggatggtg
     5161 actgttatat ctatgaaatt cgatttagag gtacgaactt tcctgccaat ggtccagtta
     5221 tgcagaagag gactgtgaaa tgggagccat ccactgagaa cttgtatgtg cgtgatggag
     5281 tgctgaaggg tgatgttgtc atggctctgt cgcttgaagg aggtggccat taccgatgtg
     5341 acttcaaaac tacttataaa gctaagaagg ttgtccagtt gccagactat cactttgtgg
     5401 accaccacat tgagattaaa agccacgaca aagattacag taatgttaat ctgcatgagc
     5461 acgccgaagc gcattctgag ctgccgaggc aggccaagcg tacgaatgat gtcgatgacc
     5521 agttgaagag gatcttccga ctgctgggga cgcccaccga ggagcagtgg ccctctatga
     5581 ccaagctgcc agactataag ccctatccga tgtacccggc cacaacatcc ctggtgaacg
     5641 tcgtgcccaa actcaatgcc acagggaggg atctgctgca gaaccttctg aagtgtaacc
     5701 ctgtccagcg tatctcagca gaagaggccc tgcagcaccc ctacttctcc gacttctgtc
     5761 cgccctgaac gcgtgatttt aaaagaaaag gggggactgg aagggctaat tcactcccac
     5821 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5881 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5941 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6001 tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7801 cgcgcacatt tccccgaaaa gtgccacctg acactagtta ttaatagtaa tcaattacgg
     7861 ggtcattagt tcatagccca tatat
//
