LOCUS       Exported                8511 bp ds-DNA     circular SYN 07-MAY-2025
DEFINITION  lentiviral vector for GATA1.
ACCESSION   .
VERSION     .
KEYWORDS    pSIN4-DDX6-Delta-303-483
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8511)
  AUTHORS   Elcheva I, Brok-Volchanskaya V, Kumar A, Liu P, Lee JH, Tong L, 
            Vodyanik M, Swanson S, Stewart R, Kyba M, Yakubov E, Cooke J, 
            Thomson JA, Slukvin I
  TITLE     Direct induction of haematoendothelial programs in human pluripotent
            stem cells by transcriptional regulators.
  JOURNAL   Nat Commun. 2014 Jul 14;5:4372. doi: 10.1038/ncomms5372.
  PUBMED    25019369
REFERENCE   2  (bases 1 to 8511)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8511
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     misc_feature    3474..3479
                     /label=EcoR1
     regulatory      3480..3488
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3489..3554
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3555..4463
                     /codon_start=1
                     /label=DDX6CDS
                     /translation="STARTENPVIMGLSSQNGQLRGPVKPTGGPGGGGTQTQQQMNQLK
                     NTNTINNGTQQQAQSMTTTIKPGDDWKKTLKLPPKDLRIKTSDVTSTKGNEFEDYCLKR
                     ELLMGIFEMGWEKPSPIQEESIPIALSGRDILARAKNGTGKSGAYLIPLLERLDLKKDN
                     IQAMVIVPTRELALQVSQICIQVSKHMGGAKVMATTGGTNLRDDIMRLDDTVHVVIATP
                     GRILDLIKKGVAKVDHVQMIVLDEADKLLSQDFVQIMEDIILTLPKNRQILLYSATFPL
                     SVQKFMNSHLQKPYEINLMEE"
     misc_feature    3555..4460
                     /label=1-302
     misc_feature    3555..3830
                     /label=IDR
     misc_feature    4029..4046
                     /label=sgDDX-1 mut
     misc_feature    4290..4292
                     /label=loss founction site
     misc_feature    4415..4420
                     /label=EcoR1
     misc_feature    4461..4463
                     /label=303-483
     misc_feature    4622..5084
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4678..4695)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4905..4924
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5117..5716
                     /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(5117..5136)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5613..5633
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5919..6152
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6303..6320)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6309..6533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      6576..6585
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        6629..6733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6734..7594
                     /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(6952..6971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7765..8353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8254..8273
                     /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 caagagcacg gccagaacag agaaccctgt tataatgggt ctgtccagtc
     3601 aaaatggtca gctgagaggc cctgtgaaac ccactggtgg ccctggagga gggggcacac
     3661 agacacagca acagatgaac cagctgaaaa acaccaacac aatcaataat ggcactcagc
     3721 agcaagcaca gagtatgacc accactatta aacctggtga tgactggaaa aagactttaa
     3781 aactccctcc aaaggatcta agaatcaaaa cttcggatgt gacctccaca aaaggaaatg
     3841 agtttgaaga ttactgtttg aaacgggagt tactgatggg aatttttgaa atgggctggg
     3901 aaaagccatc tcctattcag gaggagagca ttcccattgc tttatctggt agggatatct
     3961 tagctagagc aaaaaatgga acaggcaaga gcggtgccta cctcattccc ttacttgaac
     4021 ggctagactt aaaaaaagat aacatccaag caatggtgat tgttcccact agagaacttg
     4081 ctctacaggt cagtcaaatt tgcatccagg tcagcaaaca catgggaggg gccaaagtga
     4141 tggcaaccac aggaggaacc aatttacgag atgacataat gaggcttgat gatacagtgc
     4201 acgtggtgat tgctacccct gggagaatcc tggatcttat taagaaagga gtagcaaagg
     4261 ttgatcatgt ccagatgata gtattggatg aggcagataa gttgctgtca caggattttg
     4321 tgcagataat ggaggatatt attctcacgc tacctaaaaa caggcagatt ttactatatt
     4381 ccgctacttt ccctcttagt gtacagaagt tcatgaattc ccatttgcag aaaccctatg
     4441 agattaacct gatggaggaa taaacgcgta aactagttcg aaggatccgc atgcatctag
     4501 ggcggccaat tccgcccctc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     4561 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     4621 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     4681 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     4741 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     4801 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     4861 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     4921 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     4981 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     5041 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttgc cacaacccac
     5101 aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     5161 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     5221 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     5281 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5341 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5401 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5461 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5521 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5581 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5641 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5701 caagcccggt gcctgataat aggcggccgc tcgagaccta gaaaaacatg gagcaatcac
     5761 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     5821 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     5881 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     5941 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     6001 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6061 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6121 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6181 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac attataatag
     6241 atttagcagg aattgaacta ggagtggagc acacaggcaa agttctagag ctcgctgatc
     6301 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6361 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6421 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     6481 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     6541 ggcggaaaga accagctggg ggcgcgcccc tcgaggccgc catggtcata gctgtttgac
     6601 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6661 acattcaaat atgtatccgc tcacgagaca ataaccctga taaatgcttc aataatattg
     6721 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6781 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6841 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6901 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6961 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7021 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7081 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7141 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7201 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7261 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7321 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7381 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7441 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7501 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7561 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7621 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7741 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7801 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7861 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7921 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7981 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8041 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8101 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8161 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8221 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8281 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8341 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttctggcctt ttgcacggac
     8401 cgccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
     8461 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag t
//
