LOCUS       sequence-245282-        6799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of cDNA with blasticidin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6799)
  AUTHORS   Tarumoto Y, Lin S, Wang J, Milazzo JP, Xu Y, Lu B, Yang Z, Wei Y, 
            Polyanskaya S, Wunderlich M, Gray NS, Stegmaier K, Vakoc CR
  TITLE     Salt-Inducible Kinase inhibition suppresses acute myeloid leukemia 
            progression in vivo.
  JOURNAL   Blood. 2019 Nov 1. pii: 422697. doi: 10.1182/blood.2019001576.
  PUBMED    31697837
REFERENCE   2  (bases 1 to 6799)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Blood. 2019
            Nov 1. pii: 422697. doi: 10.1182/blood.2019001576."
COMMENT     Sequence Label: LentiV_Blast
FEATURES             Location/Qualifiers
     source          1..6799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2149..2172
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2182..2238
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2239..2637
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    2656..3244
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2709..2729)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3127..3138)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3316..3549
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3627..3748
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3664..3683)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3718..3737
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3788..3923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3943..3962)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3944..3962)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3972..3989)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3972..3988)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3981..4003)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4130..4585
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4217..4236)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4427..4448
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4611..4715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4716..4784,4785..5576)
                     /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(4934..4953)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5747..6335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6236..6255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6489..6506
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6623..6644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6659..6664,6665..6682,6683..6689)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6697..6713
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6702..6724
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6721..6737
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6721..6737
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6756..6776
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6758..6776
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga ttacaaagac
     2161 gatgacgata agggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg
     2221 gaggagaacc ccggccccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
     2281 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     2341 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     2401 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     2461 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     2521 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     2581 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaatga
     2641 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2701 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2761 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2821 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2881 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2941 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3001 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3061 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3121 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3181 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3241 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     3301 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     3361 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     3421 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     3481 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     3541 tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa
     3601 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     3661 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3721 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     3781 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3841 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3901 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     3961 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4021 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4081 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     4141 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4201 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4261 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4321 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4381 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4441 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4501 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4561 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4621 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4681 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4741 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4801 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4861 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4921 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4981 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5041 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5101 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5161 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5221 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5281 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5341 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5401 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5461 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5521 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5581 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5641 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5701 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5761 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5821 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5881 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5941 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6001 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6061 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6121 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6181 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6241 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6301 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6361 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6421 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6481 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6541 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6601 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     6661 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     6721 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     6781 aaaagctgga gctgcaagc
//
