LOCUS       Exported                9014 bp ds-DNA     circular SYN 07-MAY-2025
DEFINITION  lentiviral vector for GATA1.
ACCESSION   .
VERSION     .
KEYWORDS    pSIN4-LSM14A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9014)
  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 9014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9014
                     /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"
     primer_bind     complement(3528..3554)
                     /label=PSIN4-R
     primer_bind     3540..3575
                     /label=PSIN4-LSM14A-F
     CDS             3555..4946
                     /label=LSM14A-CDS
     primer_bind     complement(4926..4961)
                     
     primer_bind     4947..4973
                     /label=PSIN4-F
     misc_feature    5125..5587
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5181..5198)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5408..5427
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5620..6219
                     /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(5620..5639)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6116..6136
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6422..6655
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6806..6823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6812..7036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      7079..7088
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        7132..7236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7237..8097
                     /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(7455..7474)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8268..8856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8757..8776
                     /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 caagatgagc gggggcaccc cttacatcgg cagcaagatc agcctcatct
     3601 ccaaggcgga gatccgctac gagggcatcc tctacaccat cgacaccgaa aactccaccg
     3661 tagcccttgc caaagttcga tcctttggta cagaagacag accgacagat cgtccaatac
     3721 cacctcgaga tgaagtcttt gaatacatta tattccgtgg gagtgacatt aaagacctta
     3781 ctgtttgtga gccaccaaaa ccacagtgtt ctttgcctca agacccagct attgttcagt
     3841 cctcactagg ctcatcgact tcttcattcc agtccatggg ttcttatgga cctttcggca
     3901 ggatgcccac atacagtcag ttcagtccga gttccttagt tgggcagcag tttggtgctg
     3961 ttggtgttgc tggaagctct ttgacatcct ttggaacaga aacatcaaac agtggtacct
     4021 taccccaaag tagtgcggtt ggttctgcct ttacacagga tacaagatct ctaaaaacac
     4081 agttatctca aggtcgctca agccctcagt tagacccttt gagaaaaagc ccaaccatgg
     4141 aacaagcagt gcagaccgcc tcagcccact tacctgctcc agcagctgtt gggagaagga
     4201 gtcctgtatc aaccaggcct ttgccatctg ccagccaaaa ggcaggagag aatcaggagc
     4261 acaggcgagc tgaagtacac aaagtttcaa ggccagaaaa tgagcaactc agaaatgata
     4321 acaagagaca agtagctcca ggtgctcctt cagctccaag gagagggcgt gggggtcatc
     4381 ggggtggcag gggaagattt ggtattcggc gagatgggcc aatgaaattt gagaaagact
     4441 ttgactttga aagtgcaaat gcacaattca acaaggaaga gattgacaga gagtttcata
     4501 ataaacttaa attaaaagaa gataaacttg agaaacagga gaagcctgta aatggtgaag
     4561 ataaaggaga ctcaggagtt gatacccaaa acagtgaagg aaatgccgat gaagaagatc
     4621 cacttggacc taattgctat tatgacaaaa ctaaatcctt ctttgataat atttcttgtg
     4681 atgacaatag agaacggaga ccaacctggg ctgaagaaag aagattaaat gctgaaacat
     4741 ttggaatccc acttcgtcca aaccgtggcc gtgggggata cagaggcaga ggaggtcttg
     4801 gtttccgtgg tggcagaggg cgtggtggtg gcagaggtgg taccttcact gcccctcgag
     4861 gatttcgcgg tggattcaga ggaggtcgtg ggggccggga gtttgcggat tttgaatata
     4921 ggaaagacaa caaagttgct gcatagacgc gtaaactagt tcgaaggatc cgcatgcatc
     4981 tagggcggcc aattccgccc ctctcccccc cccccctctc cctccccccc cccctaacgt
     5041 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     5101 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     5161 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     5221 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     5281 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     5341 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     5401 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     5461 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     5521 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     5581 atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta caagcccacg
     5641 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
     5701 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
     5761 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     5821 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5881 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5941 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     6001 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     6061 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     6121 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     6181 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat aataggcggc cgctcgagac
     6241 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     6301 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     6361 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6421 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     6481 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6541 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6601 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6661 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6721 gagaggcctt gacattataa tagatttagc aggaattgaa ctaggagtgg agcacacagg
     6781 caaagttcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     6841 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     6901 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     6961 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     7021 gcggtgggct ctatggcttc tgaggcggaa agaaccagct gggggcgcgc ccctcgaggc
     7081 cgccatggtc atagctgttt gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7141 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcacgag acaataaccc
     7201 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7261 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7321 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7381 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7441 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7501 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7561 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7621 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7681 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7741 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7801 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7861 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7921 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7981 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8041 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8101 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8161 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8221 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8281 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8341 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8401 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8461 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8521 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8581 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8641 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8701 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8761 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8821 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8881 cggttctggc cttttgcacg gaccgccact gccaattacc tgtggtttca tttactctaa
     8941 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     9001 acaaacttag tagt
//
