LOCUS       sequence-367803-        9485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for codon-optimized 5-Lipoxygenase 
            (enzymatically inactive E376Q mutant) in mamallian cells with 
            blasticidin S resistance (bsd)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9485)
  AUTHORS   Kreiss M, Oberlis J, Seuter S, Bischoff-Kont I, Surun D, Thomas D, 
            Gobel T, Schmid T, Radmark O, Brandes RP, Furst R, Hafner AK, 
            Steinhilber D
  TITLE     Human 5-lipoxygenase regulates transcription by association to 
            euchromatin.
  JOURNAL   Biochem Pharmacol. 2022 Jul 22:115187. doi: 
            10.1016/j.bcp.2022.115187.
  PUBMED    35878796
REFERENCE   2  (bases 1 to 9485)
  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: "Biochem 
            Pharmacol. 2022 Jul 22:115187. doi: 10.1016/j.bcp.2022.115187."
COMMENT     Sequence Label: LeGO-iBSD-5LO-E376Q
FEATURES             Location/Qualifiers
     source          1..9485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(25..599,600..602,603..611)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(192..209)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     419..438
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             612..1007
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(1059..1092)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1148..1736
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1201..1221)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1619..1630)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1739..1755)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             2265..2445
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2507..3095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2587..2606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3266..4057,4058..4126))
                     /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     3889..3908
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4127..4231)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4306..4325)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4497..4876
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4878..5076
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5031..5051
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5094..5274
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5321..5446
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5943..6176
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6361..6405
                     /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    6703..6820
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     6873..6889
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6931..6964)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7035..7441
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
ORIGIN
        1 gctacgtaaa ttccgccccc cccccccctc tccctccccc ccccctaacg ttactggccg
       61 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
      121 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
      181 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
      241 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
      301 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
      361 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
      421 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
      481 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
      541 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
      601 tggccacaac cgccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
      661 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
      721 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
      781 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
      841 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
      901 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
      961 ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaaagt aaagcggccg
     1021 gccgccagca cagtggtcga aattcgtcga gggacctaat aacttcgtat agcatacatt
     1081 atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct
     1141 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1201 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1261 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     1321 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     1381 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     1441 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1501 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     1561 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     1621 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     1681 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     1741 ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     1801 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     1861 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     1921 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     1981 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     2041 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     2101 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     2161 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     2221 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     2281 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     2341 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     2401 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
     2461 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2941 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3241 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3301 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3361 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3421 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3481 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3541 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3601 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3661 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3721 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3781 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3841 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3901 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3961 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4021 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4081 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4141 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4201 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4261 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     4321 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     4381 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     4441 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     4501 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     4561 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4621 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     4681 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     4741 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4801 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4861 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     4921 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4981 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     5041 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
     5101 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5161 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5221 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     5281 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     5341 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     5401 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     5461 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     5521 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     5581 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     5641 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     5701 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     5761 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     5821 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     5881 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     5941 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     6001 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     6061 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     6121 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     6181 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     6241 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     6301 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     6361 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     6421 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     6481 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     6541 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     6601 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     6661 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     6721 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     6781 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     6841 cagagatcca gtttggttag taccgggccc gccgaggtcg acggtatcga taagctcgct
     6901 tcacgagatt ccagcaggtc gagggaccta ataacttcgt atagcataca ttatacgaag
     6961 ttatattaag ggttccaagc ttaagcggcc ggccgctgaa agaccccacc tgtaggtttg
     7021 gcaagctagc tgcagtaacg ccattttgca aggcatggaa aaataccaaa ccaagaatag
     7081 agaagttcag atcaagggcg ggtacatgaa aatagctaac gttgggccaa acaggatatc
     7141 tgcggtgagc agtttcggcc ccggcccggg gccaagaaca gatggtcacc gcagtttcgg
     7201 ccccggcccg aggccaagaa cagatggtcc ccagatatgg cccaaccctc agcagtttct
     7261 taagacccat cagatgtttc caggctcccc caaggacctg aaatgaccct gcgccttatt
     7321 tgaattaacc aatcagcctg cttctcgctt ctgttcgcgc gcttctgctt cccgagctct
     7381 ataaaagagc tcacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg
     7441 gatccgccgc caccatgccc agctacaccg tgaccgtggc caccggctct cagtggttcg
     7501 ccggcaccga cgactacatc tacctgagcc tggtcggaag cgccggctgc agcgagaaac
     7561 atctgctgga caagcccttc tacaacgact tcgagcgggg agccgtggac tcctacgatg
     7621 tgaccgtgga cgaggaactg ggcgagatcc agctggtccg catcgagaag cggaagtact
     7681 ggctgaacga cgactggtat ctgaagtaca tcaccctgaa aaccccccac ggcgactaca
     7741 tcgagttccc ttgctaccgg tggattacag gagacgtgga agtggtgctg cgggacggca
     7801 gagccaagct ggccagggac gaccagatcc acatcctgaa gcagcaccgg cggaaagagc
     7861 tggaaacccg gcagaaacag taccgctgga tggaatggaa ccccggcttc cccctgtcca
     7921 tcgacgccaa gtgccacaag gacctgcccc gggacatcca gttcgacagc gagaagggcg
     7981 tggacttcgt gctgaactac agcaaggcca tggaaaacct gttcatcaac cggttcatgc
     8041 acatgttcca gagcagctgg aacgacttcg ccgacttcga gaagatcttc gtgaagatca
     8101 gcaacaccat cagcgagaga gtgatgaacc actggcagga agatctgatg ttcggctacc
     8161 agttcctgaa cggctgcaac cccgtgctga tccggcggtg taccgagctg cctgagaagc
     8221 tgcccgtgac caccgagatg gtggaatgca gcctggaacg gcagctgagc ctggaacagg
     8281 aagtgcagca gggcaacatc ttcatcgtgg atttcgagct gctggacggc attgacgcca
     8341 acaagaccga cccctgcacc ctgcagtttc tggccgctcc catctgcctg ctgtacaaga
     8401 acctggccaa caagatcgtg cctatcgcca tccagctgaa ccagatcccc ggcgacgaga
     8461 accccatctt cctgcccagc gacgccaaat acgactggct gctggccaag atctgggtcc
     8521 gctccagcga cttccacgtg caccagacca tcacccatct gctgcggacc cacctggtgt
     8581 cccaggtgtt cggaatcgcc atgtacagac agctgcccgc cgtgcaccct atcttcaagc
     8641 tgctggtggc ccacgtgcgg ttcacaatcg ccatcaacac caaggccaga gagcagctga
     8701 tctgcgagtg cggcctgttc gacaaggcca acgccacagg cggcggaggc cacgtgcaga
     8761 tggtgcagag agccatgaag gacctgacct acgccagcct gtgcttcccc gaggccatca
     8821 aggccagggg catggaaagc aaagaggaca tcccctacta cttctaccgg gacgacggcc
     8881 tgctggtctg ggaggccatc agaaccttca ccgccgaggt ggtggacatc tactacgagg
     8941 gcgaccaggt ggtggaagag gaccccgagc tgcaggactt cgtgaacgac gtgtacgtgt
     9001 acggcatgcg gggcagaaag agcagcggct tccccaagag cgtgaagtcc cgcgagcagc
     9061 tgagcgagta cctgaccgtg gtgatcttca ccgccagcgc ccagcacgcc gccgtgaact
     9121 tcggccagta cgactggtgc agctggatcc ccaacgcccc tcccaccatg agagcccccc
     9181 cacctacagc caagggcgtg gtgacaatcg agcagatcgt ggacaccctg cccgaccggg
     9241 gcagaagctg ttggcatctg ggagccgtgt gggccctgag ccagttccag gaaaacgagc
     9301 tgttcctggg catgtacccc gaggaacact ttatcgagaa gcccgtgaaa gaagccatgg
     9361 cccggttccg gaagaacctg gaagccatcg tgtccgtgat cgccgagcgg aacaagaaga
     9421 agcagctgcc ctactactac ctgtcccccg acagaatccc taacagcgtg gccatctgag
     9481 cggcc
//
