LOCUS       sequence-479093-        9281 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  Expresses human DDX6-DDX3X-IDR1+DDX6-Delta-IDR fused to 3X Flag in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9281)
  TITLE     Su lab plasmids 2025
REFERENCE   2  (bases 1 to 9281)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pSIN4-DDX3X-IDR1+DDX6-Delta-IDR
FEATURES             Location/Qualifiers
     source          1..9281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1299..1532
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1717..1761
                     /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    2048..2165
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2294..3472
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2525..3463
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3420..3440
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3480..3489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3489..3554
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    5283..5854
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5448..5465)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5675..5694
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5887..6486
                     /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(5887..5906)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6383..6403
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6689..6922
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7073..7090)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7079..7303
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      7346..7355
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        7399..7503
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7504..7572,7573..8364)
                     /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     complement(7722..7741)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8535..9123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9024..9043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1201 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1261 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1321 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1381 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1441 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1501 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1561 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1621 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1681 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1741 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1801 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     1861 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     1921 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     1981 gacagatcca ttcgattagt gaacggatct cgacggtatc gccacaaatg gcagtattca
     2041 tccacaattt taaaagaaag ggggggattg gggggtacag tgcaggggaa agaatagtag
     2101 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2161 attttcgggt ttattacagg gacagcagag atccactttg gatcgataag ctttgcaaag
     2221 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
     2281 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     2341 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
     2401 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     2461 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     2521 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
     2581 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
     2641 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
     2701 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
     2761 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
     2821 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     2881 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     2941 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     3001 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     3061 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     3121 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     3181 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     3241 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     3301 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     3361 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     3421 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattcg
     3481 ccaccatgga ctacaaggac cacgacggtg actacaagga ccacgacatc gactacaagg
     3541 acgacgacga caagagtcat gtggcagtgg aaaatgcgct cgggctggac cagcagtttg
     3601 ctggcctaga cctgaactct tcagataatc agagtggagg aagtacagcc agcaaagggc
     3661 gctatattcc tcctcattta aggaaccgag aagctactaa aggtttctac gataaagaca
     3721 gttcagggtg gagttctagc aaagataagg atgcgtatag cagttttgga tctcgtagtg
     3781 attcaagagg gaagtctagc ttcttcagtg atcgtggaag tggatcaagg ggaaggtttg
     3841 atgatcgtgg acggagtgat tacgatggca ttggcagccg tggtgacaga agtggctttg
     3901 gcaaatttga acgtggtgga aacagtcgct ggtgtgacaa atcagatgaa gatgattggt
     3961 caaaaccact cccaccaagt gaacgcttgg aacaggaact cttttctgga ggcaacactg
     4021 ggattaattt tgagaaatac gatgacattc cagttgagaa aggaaatgag tttgaagatt
     4081 actgtttgaa acgggagtta ctgatgggaa tttttgaaat gggctgggaa aagccatctc
     4141 ctattcagga ggagagcatt cccattgctt tatctggtag ggatatctta gctagagcaa
     4201 aaaatggaac aggcaagagc ggtgcctacc tcattccctt acttgaacgg ctagacttaa
     4261 aaaaagataa catccaagca atggtgattg ttcccactag agaacttgct ctacaggtca
     4321 gtcaaatttg catccaggtc agcaaacaca tgggaggggc caaagtgatg gcaaccacag
     4381 gaggaaccaa tttacgagat gacataatga ggcttgatga tacagtgcac gtggtgattg
     4441 ctacccctgg gagaatcctg gatcttatta agaaaggagt agcaaaggtt gatcatgtcc
     4501 agatgatagt attggatgag gcagataagt tgctgtcaca ggattttgtg cagataatgg
     4561 aggatattat tctcacgcta cctaaaaaca ggcagatttt actatattcc gctactttcc
     4621 ctcttagtgt acagaagttc atgaattccc atttgcagaa accctatgag attaacctga
     4681 tggaggaact aactctgaag ggagtaaccc agtactacgc atatgtaact gagcgccaaa
     4741 aagtacactg cctcaacaca cttttctcca ggcttcagat aaaccagtcg atcattttct
     4801 gtaactcctc tcagcgagtt gaattgctag ccaagaagat ttctcaactg ggttattctt
     4861 gcttctatat tcatgctaaa atgaggcagg aacatcgaaa tcgtgtattt catgatttcc
     4921 gaaatggctt atgccgcaat cttgtttgca ctgatctgtt tacccgaggt attgatatac
     4981 aagctgtgaa tgtggtaata aactttgatt tcccaaagct ggcagagacc tatctccatc
     5041 gtattggaag atcaggtcgc tttggtcatc ttggcttagc catcaacttg atcacatatg
     5101 atgatcgctt caacctgaaa agtattgagg agcagctggg aacagaaatt aaacctattc
     5161 cgagcaacat tgataagagc ctgtatgtgg cagaatacca cagcgagcct gtagaagatg
     5221 agaaacctta aacgcgtaaa ctagttcgaa ggatccgcat gcatctaggg cggccaattc
     5281 cgcccctctc cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     5341 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     5401 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     5461 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     5521 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     5581 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     5641 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     5701 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     5761 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     5821 ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga
     5881 ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg
     5941 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac
     6001 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     6061 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     6121 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     6181 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     6241 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     6301 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     6361 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     6421 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     6481 gcctgataat aggcggccgc tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
     6541 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
     6601 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     6661 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     6721 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     6781 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6841 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6901 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     6961 cttgcaaaga aatgaatatc agagagtgag aggccttgac attataatag atttagcagg
     7021 aattgaacta ggagtggagc acacaggcaa agttctagag ctcgctgatc agcctcgact
     7081 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     7141 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     7201 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     7261 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     7321 accagctggg ggcgcgcccc tcgaggccgc catggtcata gctgtttgac gtcaggtggc
     7381 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7441 atgtatccgc tcacgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7501 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7561 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7621 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7681 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7741 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7801 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7861 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7921 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7981 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     8041 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     8101 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     8161 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8221 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8281 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8341 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8401 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8461 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8521 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8581 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8641 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8701 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8761 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8821 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8881 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8941 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     9001 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     9061 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     9121 aaaaacgcca gcaacgcggc ctttttacgg ttctggcctt ttgcacggac cgccactgcc
     9181 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     9241 agaaattgta tttgttaaat atgtactaca aacttagtag t
//
