LOCUS       sequence-234684-        8307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgGAL4 control. Lentiviral mouse U6 (mU6) promoter-driven expression
            vector that coexpessed Puro-T2A-mCherry from a CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8307)
  AUTHORS   Lee JY, Chang JK, Dominguez AA, Lee HP, Nam S, Chang J, Varma S, Qi 
            LS, West RB, Chaudhuri O
  TITLE     YAP-independent mechanotransduction drives breast cancer 
            progression.
  JOURNAL   Nat Commun. 2019 Apr 23;10(1):1848. doi: 10.1038/s41467-019-09755-0.
  PUBMED    31015465
REFERENCE   2  (bases 1 to 8307)
  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: "Nat 
            Commun."; date: "2019-04-23"; volume: "10(1):1848. doi"; pages: " 
            10.1038/s41467-019-09755-0"
COMMENT     Sequence Label: pSLQ1371-sgGAL4
FEATURES             Location/Qualifiers
     source          1..8307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..211
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             229..409
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    456..581
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1838..1955
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2208..2241)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2245..2265
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     enhancer        2558..2861
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2862..3065
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3099..3695
                     /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(3099..3118)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3595..3615
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3705..3758
                     /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      3765..3774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3771..4481
                     /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(3916..3934)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4184..4203
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(4501..4534)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4590..5178
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4643..4663)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5061..5072)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5181..5197)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5182..5198)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5707..5887
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5949..6537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6029..6048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6708..7499,7500..7568))
                     /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     7331..7350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7569..7673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7748..7767)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7939..8307,1..11)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
       61 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      121 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      181 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
      241 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      301 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      361 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      421 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      481 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      541 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      601 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      661 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      721 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      781 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
      841 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
      901 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc
     1801 gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     1861 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     1921 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     1981 atccagtttg gttagtaccg ggcccgctct agagatccga cgcgccatct ctaggcccgc
     2041 gccggccccc tcgcacggac ttgtgggaga agctcggcta ctcccctgcc ccggttaatt
     2101 tgcatataat atttcctagt aactatagag gcttaatgtg cgataaaaga cagataatct
     2161 gttcttttta atactagcta cattttacat gataggcttg gatttctata acttcgtata
     2221 gcatacatta tacgaagtta taaacagcac aaaaggaaac tcaccctaac tgtaaagtaa
     2281 ttgtgtgttt tgagactata agtatccctt ggagaaccac cttgttggag cactgtcctc
     2341 cgaacgtgtt taagagctat gctggaaaca gcatagcaag tttaaataag gctagtccgt
     2401 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctcgagtact aggatccatt
     2461 aggcggccgc gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     2521 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2581 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2641 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2701 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2761 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2821 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2881 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     2941 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     3001 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     3061 gagctggttt agtgaaccgt cagatccgct agcccaccat gaccgagtac aagcccacgg
     3121 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     3181 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     3241 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     3301 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     3361 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     3421 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     3481 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     3541 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     3601 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     3661 cgcgcacctg gtgcatgacc cgcaagcccg gtgccggatc gggagagggc agaggaagtc
     3721 tgctaacatg cggtgacgtc gaggagaatc ctggcccacc ggtcgccacc atggtgagca
     3781 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     3841 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     3901 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     3961 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     4021 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     4081 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     4141 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     4201 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     4261 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     4321 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     4381 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     4441 gccgccactc caccggcggc atggacgagc tgtacaagta agaattcgtc gagggaccta
     4501 ataacttcgt atagcataca ttatacgaag ttatacatgt ttaagggttc cggttccact
     4561 aggtacaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     4621 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4681 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4741 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4801 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4861 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4921 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4981 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5041 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5101 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     5161 tttgggccgc ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga
     5221 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
     5281 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
     5341 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     5401 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     5461 cctgattggc agaactacac accagggcca gggatcagat atccactgac ctttggatgg
     5521 tgctacaagc tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac
     5581 acccgcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta
     5641 gagtggaggt ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac
     5701 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5761 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5821 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5881 tctagcagca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5941 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6001 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6061 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6121 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6181 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6241 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6301 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6361 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6421 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6481 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6541 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6601 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6661 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6721 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6781 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6841 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6901 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6961 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7021 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7081 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7141 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7201 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7261 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7321 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7381 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7441 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7501 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7561 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7621 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7681 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     7741 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     7801 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7861 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     7921 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     7981 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     8041 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     8101 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     8161 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     8221 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     8281 ggcagtacat ctacgtatta gtcatcg
//
