LOCUS       sequence-364559-        8308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgGal4-expressing vector modified from Addgene plasmid #51024.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8308)
  AUTHORS   Liu X, Zhang Y, Chen Y, Li M, Zhou F, Li K, Cao H, Ni M, Liu Y, Gu 
            Z, Dickerson KE, Xie S, Hon GC, Xuan Z, Zhang MQ, Shao Z, Xu J
  TITLE     In Situ Capture of Chromatin Interactions by Biotinylated dCas9.
  JOURNAL   Cell. 2017 Aug 24;170(5):1028-1043.e19. doi: 
            10.1016/j.cell.2017.08.003.
  PUBMED    28841410
REFERENCE   2  (bases 1 to 8308)
  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: 
            "10.1016/j.cell.2017.08"; journalName: "Cell"; date: "2017-08-24- 
            24"; volume: "170"; issue: "5"; pages: "1028-1043.e19"
COMMENT     Sequence Label: pSLQ1651-sgRNA(F+E)-sgGal4
FEATURES             Location/Qualifiers
     source          1..8308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..250
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             284..880
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(284..303)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     780..800
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             890..943
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      950..959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             956..1666
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1101..1119)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1369..1388
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1686..1719)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1775..2363
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1828..1848)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2246..2257)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2366..2382)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2367..2383)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2892..3072
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3134..3722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3214..3233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3893..4684,4685..4753))
                     /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     4516..4535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4754..4858)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4933..4952)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5124..5503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5505..5703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5721..5901
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5948..6073
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6570..6803
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6988..7032
                     /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    7330..7447
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(7700..7733)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     7737..7757
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     enhancer        join(8051..8308,1..46)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
       61 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      121 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      181 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      241 aagcagagct ggtttagtga accgtcagat ccgctagccc accatgaccg agtacaagcc
      301 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
      361 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
      421 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
      481 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
      541 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
      601 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
      661 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
      721 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
      781 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
      841 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc ggatcgggag agggcagagg
      901 aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaccggtcg ccaccatggt
      961 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     1021 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1081 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1141 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     1201 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1261 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1321 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1381 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1441 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1501 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     1561 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     1621 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagaat tcgtcgaggg
     1681 acctaataac ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt
     1741 ccactaggta caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     1801 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1861 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1921 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1981 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2041 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2101 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2161 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     2221 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2281 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2341 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac
     2401 atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag
     2461 cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga
     2521 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     2581 taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
     2641 acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg
     2701 gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag
     2761 agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
     2821 tattagagtg gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc
     2881 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     2941 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     3001 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     3061 aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3241 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3301 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3361 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3421 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3481 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3541 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3601 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3661 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3721 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3781 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3841 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3961 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4021 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4081 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4321 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4381 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4561 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4741 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4801 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4861 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     4921 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     4981 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     5041 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     5101 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     5161 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     5221 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     5281 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     5341 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     5401 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     5461 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     5521 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     5581 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     5641 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     5701 cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5761 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5821 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5881 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     5941 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     6001 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     6061 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     6121 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     6181 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     6241 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     6301 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     6361 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg
     6421 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     6481 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     6541 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     6601 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     6661 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     6721 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     6781 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     6841 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     6901 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     6961 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     7021 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     7081 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     7141 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     7201 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     7261 attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt
     7321 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     7381 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     7441 aaattttcgg gtttattaca gggacagcag agatccagtt tggttagtac cgggcccgct
     7501 ctagagatcc gacgcgccat ctctaggccc gcgccggccc cctcgcacgg acttgtggga
     7561 gaagctcggc tactcccctg ccccggttaa tttgcatata atatttccta gtaactatag
     7621 aggcttaatg tgcgataaaa gacagataat ctgttctttt taatactagc tacattttac
     7681 atgataggct tggatttcta taacttcgta tagcatacat tatacgaagt tataaacagc
     7741 acaaaaggaa actcacccta actgtaaagt aattgtgtgt tttgagacta taagtatccc
     7801 ttggagaacc accttgttgg aacgactagt taggcgtgta gtttaagagc tatgctggaa
     7861 acagcatagc aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     7921 cggtgctttt tttctcgagt actaggatcc attaggcggc cgcgtggata accgtattac
     7981 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     8041 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     8101 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     8161 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     8221 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     8281 atgcccagta catgacctta tgggactt
//
