LOCUS       Exported                8735 bp ds-DNA     circular SYN 07-MAY-2025
DEFINITION  lentiviral vector for GATA1.
ACCESSION   .
VERSION     .
KEYWORDS    pSIN4-BCAT1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8735)
  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 8735)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 15, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8735
                     /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..4643
                     /codon_start=1
                     /label=BCAT1
                     /translation="GLQ*RMLRRVYRRRRIKRGGGDF*G*RPNSHTSYHFKGKTRPQ*S
                     GFWNCVHGSYADGGVVLRVWMGETSYQASSEPVIAPWLISFALCSGII*RIEGISRSR*
                     *NSTVSAKPQHG*NVSLCCEGNSAGI*QRRALRVYSTACEIGSRMGPIFNIC*SVYSSY
                     IHWN*AFSWSQEAYQSPALCTLEPSGTLFFKWNL*SSVPVGQSQVCKSLERWNWGLQDG
                     RELRLISFCPM*SSR*WVSAGPVALWRGPSDH*SGNYESFSLLDK*RWRRRTGNSSTRW
                     HHSSRSDKAVHSGPGTSVG*I*GVREIPHHG*LDNSPGGEQSERDVWLWYSLCCLPSF*
                     YTVQRRDNTHSNYGEWS*AGKPHLEQIN*YPVWKRRERLDNCAI"
     misc_feature    3614..3638
                     /label=0.65/0.93
     misc_feature    3639..3663
                     /label=0.5/0.97
     misc_feature    3789..3813
                     /label=0.75/0.94
     primer_bind     4025..4045
                     /label=hBCAT1-F-1
     misc_feature    4086..4109
                     /label=0.65/0.8
     primer_bind     complement(4106..4127)
                     /label=hBCAT1-R-1
     primer_bind     4359..4379
                     /label=hBCAT1-F-2
     misc_feature    4387..4411
                     /label=0.55/0.96
     misc_feature    4389..4413
                     /label=0.5/0.97
     primer_bind     complement(4432..4452)
                     /label=hBCAT1-R-2
     misc_feature    4539..4563
                     /label=0.5/0.99
     CDS             4644..4667
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4846..5308
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4902..4919)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5129..5148
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5341..5940
                     /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(5341..5360)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5837..5857
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6143..6376
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6527..6544)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6533..6757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      6800..6809
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        6853..6957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6958..7818
                     /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(7176..7195)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7989..8577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8478..8497
                     /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 ccaccatgaa ggattgcagt aacggatgct ccgcagagtg taccggagaa ggaggatcaa
     3541 aagaggtggt ggggactttt aaggctaaag acctaatagt cacaccagct accattttaa
     3601 aggaaaaacc agaccccaat aatctggttt ttggaactgt gttcacggat catatgctga
     3661 cggtggagtg gtcctcagag tttggatggg agaaacctca tatcaagcct cttcagaacc
     3721 tgtcattgca ccctggctca tcagctttgc actatgcagt ggaattattt gaaggattga
     3781 aggcatttcg aggagtagat aataaaattc gactgtttca gccaaacctc aacatggata
     3841 gaatgtatcg ctctgctgtg agggcaactc tgccggtatt tgacaaagaa gagctcttag
     3901 agtgtattca acagcttgtg aaattggatc aagaatgggt cccatattca acatctgcta
     3961 gtctgtatat tcgtcctaca ttcattggaa ctgagccttc tcttggagtc aagaagccta
     4021 ccaaagccct gctctttgta ctcttgagcc cagtgggacc ttatttttca agtggaacct
     4081 ttaatccagt gtccctgtgg gccaatccca agtatgtaag agcctggaaa ggtggaactg
     4141 gggactgcaa gatgggaggg aattacggct catctctttt tgcccaatgt gaagcagtag
     4201 ataatgggtg tcagcaggtc ctgtggctct atggagagga ccatcagatc actgaagtgg
     4261 gaactatgaa tctttttctt tactggataa atgaagatgg agaagaagaa ctggcaactc
     4321 ctccactaga tggcatcatt cttccaggag tgacaaggcg gtgcattctg gacctggcac
     4381 atcagtgggg tgaatttaag gtgtcagaga gatacctcac catggatgac ttgacaacag
     4441 ccctggaggg gaacagagtg agagagatgt ttggctctgg tacagcctgt gttgtttgcc
     4501 cagtttctga tatactgtac aaaggcgaga caatacacat tccaactatg gagaatggtc
     4561 ctaagctggc aagccgcatc ttgagcaaat taactgatat ccagtatgga agagaagaga
     4621 gcgactggac aattgtgcta tccgactaca aagacgatga cgacaagtga cgcgtaaact
     4681 agttcgaagg atccgcatgc atctagggcg gccaattccg cccctctccc cccccccccc
     4741 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     4801 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     4861 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     4921 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     4981 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     5041 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     5101 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     5161 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     5221 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     5281 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccacaaggag acgaccttcc
     5341 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     5401 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     5461 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5521 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5581 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5641 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5701 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5761 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5821 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5881 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5941 taataggcgg ccgctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     6001 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     6061 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6121 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     6181 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6241 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6301 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6361 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6421 aagaaatgaa tatcagagag tgagaggcct tgacattata atagatttag caggaattga
     6481 actaggagtg gagcacacag gcaaagttct agagctcgct gatcagcctc gactgtgcct
     6541 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6601 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6661 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     6721 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     6781 tgggggcgcg cccctcgagg ccgccatggt catagctgtt tgacgtcagg tggcactttt
     6841 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6901 ccgctcacga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6961 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7021 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7081 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7141 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7201 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7261 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     7321 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     7381 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     7441 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     7501 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7561 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7621 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7681 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7741 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7801 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7861 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7921 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7981 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8041 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     8101 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     8161 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     8221 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     8281 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     8341 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     8401 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     8461 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     8521 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8581 gccagcaacg cggccttttt acggttctgg ccttttgcac ggaccgccac tgccaattac
     8641 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     8701 tgtatttgtt aaatatgtac tacaaactta gtagt
//
