LOCUS       sequence-199002-        7633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid encoding TRE3G sgRNA (for SpCas9) and mTagBFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7633)
  AUTHORS   Zhou XX, Zou X, Chung HK, Gao Y, Liu Y, Qi LS, Lin MZ
  TITLE     A Single-Chain Photoswitchable CRISPR-Cas9 Architecture for 
            Light-Inducible Gene Editing and Transcription.
  JOURNAL   ACS Chem Biol. 2017 Sep 29. doi: 10.1021/acschembio.7b00603.
  PUBMED    28938067
REFERENCE   2  (bases 1 to 7633)
  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: "ACS Chem 
            Biol. 2017 Sep 29. doi: 10.1021/acschembio.7b00603."
COMMENT     Sequence Label: U6-spTRE3G-CMV-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..7633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1447..1680
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1865..1909
                     /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    2207..2324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2577..2610)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2614..2634
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     enhancer        2928..3231
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3232..3435
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    complement(4196..4229)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4285..4873
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4338..4358)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4756..4767)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4876..4892)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4877..4893)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5402..5582
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5644..6232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5724..5743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6403..7194,7195..7263))
                     /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     7026..7045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7264..7368)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7443..7462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     1321 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1381 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     1441 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     1501 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     1561 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     1621 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     1681 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     1741 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     1801 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     1861 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     1921 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     1981 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2041 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2101 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2161 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2341 acagcagaga tccagtttgg ttagtaccgg gcccgctcta gagatccgac gcgccatctc
     2401 taggcccgcg ccggccccct cgcacggact tgtgggagaa gctcggctac tcccctgccc
     2461 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2521 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2581 cttcgtatag catacattat acgaagttat aaacagcaca aaaggaaact caccctaact
     2641 gtaaagtaat tgtgtgtttt gagactataa gtatcccttg gagaaccacc ttgttggtac
     2701 gttctctatc actgatagtt taagagctat gctggaaaca gcatagcaag tttaaataag
     2761 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctcgagtact
     2821 aggatccatt aggcggccgc gtggataacc gtattaccgc catgcattag ttattaatag
     2881 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2941 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3001 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3061 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     3121 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3181 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3241 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3301 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3361 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3421 tatataagca gagctggttt agtgaaccgt cagatccgct agcccaccat ggtgtctaag
     3481 ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     3541 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     3601 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     3661 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     3721 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     3781 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     3841 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     3901 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     3961 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     4021 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     4081 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     4141 tactgcgacc tccctagcaa actggggcac aaataagaat tcgtcgaggg acctaataac
     4201 ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt ccactaggta
     4261 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     4321 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4381 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4441 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4501 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4561 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4621 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4681 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4741 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4801 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4861 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
     4921 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     4981 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     5041 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     5101 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     5161 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     5221 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     5281 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     5341 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     5401 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5461 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     5521 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     5581 cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5641 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5701 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5761 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5821 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5881 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5941 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6001 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6061 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6121 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6181 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6241 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6301 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6361 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6421 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6481 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6541 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6601 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6661 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6721 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6781 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6841 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6901 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6961 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7021 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7081 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7141 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7201 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7261 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7321 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7381 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     7441 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7501 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     7561 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     7621 agatatacgc gtt
//
