LOCUS       sequence-234681-        8308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgSTAT3-1 sequence: GTCAGGATAGAGATAGACCAG. 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 8308)
  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 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"; journalName: "Nat 
            Commun."; date: "2019-04-23"; volume: "10(1):1848. doi"; pages: " 
            10.1038/s41467-019-09755-0"
COMMENT     Sequence Label: pSLQ1371-sgSTAT3-1
FEATURES             Location/Qualifiers
     source          1..8308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             600..1196
                     /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(600..619)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1096..1116
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1206..1259
                     /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      1266..1275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1272..1982
                     /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(1417..1435)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1685..1704
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2002..2035)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2091..2679
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2144..2164)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2562..2573)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2682..2698)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2683..2699)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3208..3388
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3450..4038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3530..3549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4209..5000,5001..5069))
                     /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     4832..4851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5070..5174)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5249..5268)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5440..5819
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5821..6019
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6037..6217
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6264..6389
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6886..7119
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7304..7348
                     /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    7646..7763
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(8016..8049)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     8053..8073
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcccacca
      601 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
      661 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
      721 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
      781 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
      841 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
      901 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
      961 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     1021 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     1081 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     1141 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggat
     1201 cgggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccac
     1261 cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     1321 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     1381 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     1441 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     1501 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     1561 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     1621 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1681 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1741 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1801 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1861 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1921 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     1981 aagaattcgt cgagggacct aataacttcg tatagcatac attatacgaa gttatacatg
     2041 tttaagggtt ccggttccac taggtacaat tcgatatcaa gcttatcgat aatcaacctc
     2101 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     2161 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     2221 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     2281 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     2341 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     2401 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     2461 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     2521 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     2581 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2641 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgat
     2701 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     2761 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
     2821 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     2881 gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
     2941 taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga
     3001 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa
     3061 gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat
     3121 gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg
     3181 gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg
     3241 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     3301 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     3361 ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg
     3421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3961 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4021 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4081 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4141 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4201 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4261 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4321 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4381 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4441 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4501 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4561 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4621 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4681 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4741 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4801 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4861 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4921 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4981 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5041 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5101 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5161 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     5221 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5281 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     5341 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     5401 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     5461 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     5521 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     5581 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     5641 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     5701 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     5761 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     5821 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     5881 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     5941 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     6001 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
     6061 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6121 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6181 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     6241 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     6301 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     6361 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     6421 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     6481 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     6541 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     6601 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     6661 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     6721 agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     6781 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     6841 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     6901 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     6961 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     7021 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     7081 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     7141 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     7201 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     7261 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     7321 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     7381 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     7441 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     7501 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     7561 agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca
     7621 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     7681 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     7741 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt
     7801 tagtaccggg cccgctctag agatccgacg cgccatctct aggcccgcgc cggccccctc
     7861 gcacggactt gtgggagaag ctcggctact cccctgcccc ggttaatttg catataatat
     7921 ttcctagtaa ctatagaggc ttaatgtgcg ataaaagaca gataatctgt tctttttaat
     7981 actagctaca ttttacatga taggcttgga tttctataac ttcgtatagc atacattata
     8041 cgaagttata aacagcacaa aaggaaactc accctaactg taaagtaatt gtgtgttttg
     8101 agactataag tatcccttgg agaaccacct tgttggtcag gatagagata gaccaggttt
     8161 aagagctatg ctggaaacag catagcaagt ttaaataagg ctagtccgtt atcaacttga
     8221 aaaagtggca ccgagtcggt gctttttttc tcgagtacta ggatccatta ggcggccgcg
     8281 tggataaccg tattaccgcc atgcatta
//
