LOCUS       sequence-479083-        8794 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  Expresses human 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 8794)
  TITLE     Su lab plasmids 2025
REFERENCE   2  (bases 1 to 8794)
  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-DDX6-Delta-IDR
FEATURES             Location/Qualifiers
     source          1..8794
                     /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    join(4794..5368,5369..5371,5372..5380)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4961..4978)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5188..5207
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5400..5999
                     /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(5400..5419)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5896..5916
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6202..6435
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6586..6603)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6592..6816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      6859..6868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        6912..7016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7017..7085,7086..7877)
                     /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(7235..7254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8048..8636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8537..8556
                     /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 caagaaagga aatgagtttg aagattactg tttgaaacgg gagttactga
     3601 tgggaatttt tgaaatgggc tgggaaaagc catctcctat tcaggaggag agcattccca
     3661 ttgctttatc tggtagggat atcttagcta gagcaaaaaa tggaacaggc aagagcggtg
     3721 cctacctcat tcccttactt gaacggctag acttaaaaaa agataacatc caagcaatgg
     3781 tgattgttcc cactagagaa cttgctctac aggtcagtca aatttgcatc caggtcagca
     3841 aacacatggg aggggccaaa gtgatggcaa ccacaggagg aaccaattta cgagatgaca
     3901 taatgaggct tgatgataca gtgcacgtgg tgattgctac ccctgggaga atcctggatc
     3961 ttattaagaa aggagtagca aaggttgatc atgtccagat gatagtattg gatgaggcag
     4021 ataagttgct gtcacaggat tttgtgcaga taatggagga tattattctc acgctaccta
     4081 aaaacaggca gattttacta tattccgcta ctttccctct tagtgtacag aagttcatga
     4141 attcccattt gcagaaaccc tatgagatta acctgatgga ggaactaact ctgaagggag
     4201 taacccagta ctacgcatat gtaactgagc gccaaaaagt acactgcctc aacacacttt
     4261 tctccaggct tcagataaac cagtcgatca ttttctgtaa ctcctctcag cgagttgaat
     4321 tgctagccaa gaagatttct caactgggtt attcttgctt ctatattcat gctaaaatga
     4381 ggcaggaaca tcgaaatcgt gtatttcatg atttccgaaa tggcttatgc cgcaatcttg
     4441 tttgcactga tctgtttacc cgaggtattg atatacaagc tgtgaatgtg gtaataaact
     4501 ttgatttccc aaagctggca gagacctatc tccatcgtat tggaagatca ggtcgctttg
     4561 gtcatcttgg cttagccatc aacttgatca catatgatga tcgcttcaac ctgaaaagta
     4621 ttgaggagca gctgggaaca gaaattaaac ctattccgag caacattgat aagagcctgt
     4681 atgtggcaga ataccacagc gagcctgtag aagatgagaa accttaaacg cgtaaactag
     4741 ttcgaaggat ccgcatgcat ctagggcggc caattccgcc cctctccccc ccccccctct
     4801 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     4861 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     4921 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     4981 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     5041 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     5101 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     5161 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     5221 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     5281 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     5341 gttttccttt gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca
     5401 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     5461 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     5521 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5581 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5641 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5701 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5761 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5821 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5881 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5941 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat
     6001 aataggcggc cgctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     6061 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     6121 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     6181 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     6241 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6301 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6361 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6421 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     6481 agaaatgaat atcagagagt gagaggcctt gacattataa tagatttagc aggaattgaa
     6541 ctaggagtgg agcacacagg caaagttcta gagctcgctg atcagcctcg actgtgcctt
     6601 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6661 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6721 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6781 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     6841 gggggcgcgc ccctcgaggc cgccatggtc atagctgttt gacgtcaggt ggcacttttc
     6901 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6961 cgctcacgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     7021 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     7081 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7141 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7201 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7261 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7321 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7381 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7441 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7501 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7561 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7621 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7681 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7741 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7801 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7861 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7921 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7981 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8161 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8641 ccagcaacgc ggccttttta cggttctggc cttttgcacg gaccgccact gccaattacc
     8701 tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt
     8761 gtatttgtta aatatgtact acaaacttag tagt
//
