LOCUS       sequence-360423-        9484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for codon-optimized 5-Lipoxygenase (WT)
            in mamallian cells with blasticidin S resistance (bsd)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9484)
  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 9484)
  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-WT
FEATURES             Location/Qualifiers
     source          1..9484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(24..598,599..601,602..610)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(191..208)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     418..437
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             611..1006
                     /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(1058..1091)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1147..1735
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1200..1220)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1618..1629)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1738..1754)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             2264..2444
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2506..3094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2586..2605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3265..4056,4057..4125))
                     /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     3888..3907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4126..4230)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4305..4324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4496..4875
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4877..5075
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5030..5050
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5093..5273
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5320..5445
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5942..6175
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6360..6404
                     /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    6702..6819
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     6872..6888
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6930..6963)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7034..7440
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
ORIGIN
        1 gctacgtaaa ttccgccccc ccccccctct ccctcccccc cccctaacgt tactggccga
       61 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
      121 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
      181 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
      241 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
      301 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
      361 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
      421 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
      481 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
      541 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
      601 ggccacaacc gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
      661 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
      721 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
      781 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
      841 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
      901 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
      961 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaaagta aagcggccgg
     1021 ccgccagcac agtggtcgaa attcgtcgag ggacctaata acttcgtata gcatacatta
     1081 tacgaagtta tacatgttta agggttccgg ttccactagg tacaattcga tatcaagctt
     1141 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     1201 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     1261 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     1321 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     1381 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     1441 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     1501 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     1561 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     1621 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     1681 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     1741 taccgtcgac ctcgatcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     1801 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     1861 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     1921 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     1981 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc
     2041 agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     2101 gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag
     2161 cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct
     2221 agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga
     2281 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     2341 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     2401 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatgt gagcaaaagg
     2461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2701 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2941 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3121 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3181 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3241 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3301 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3361 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3421 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3481 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3541 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3601 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3661 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3721 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3781 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3841 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3901 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3961 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4021 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4081 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4141 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4261 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     4321 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     4381 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     4441 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
     4501 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     4561 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4621 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4681 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4741 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4801 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4861 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4921 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4981 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     5041 ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct
     5101 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     5161 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     5221 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
     5281 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
     5341 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
     5401 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
     5461 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
     5521 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
     5581 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
     5641 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
     5701 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
     5761 agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata
     5821 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     5881 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     5941 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     6001 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     6061 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     6121 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     6181 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     6241 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     6301 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     6361 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     6421 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     6481 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     6541 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     6601 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
     6661 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     6721 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     6781 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     6841 agagatccag tttggttagt accgggcccg ccgaggtcga cggtatcgat aagctcgctt
     6901 cacgagattc cagcaggtcg agggacctaa taacttcgta tagcatacat tatacgaagt
     6961 tatattaagg gttccaagct taagcggccg gccgctgaaa gaccccacct gtaggtttgg
     7021 caagctagct gcagtaacgc cattttgcaa ggcatggaaa aataccaaac caagaataga
     7081 gaagttcaga tcaagggcgg gtacatgaaa atagctaacg ttgggccaaa caggatatct
     7141 gcggtgagca gtttcggccc cggcccgggg ccaagaacag atggtcaccg cagtttcggc
     7201 cccggcccga ggccaagaac agatggtccc cagatatggc ccaaccctca gcagtttctt
     7261 aagacccatc agatgtttcc aggctccccc aaggacctga aatgaccctg cgccttattt
     7321 gaattaacca atcagcctgc ttctcgcttc tgttcgcgcg cttctgcttc ccgagctcta
     7381 taaaagagct cacaacccct cactcggcgc gccagtcctc cgattgactg agtcgcccgg
     7441 atccgccgcc accatgccca gctacaccgt gaccgtggcc accggctctc agtggttcgc
     7501 cggcaccgac gactacatct acctgagcct ggtcggaagc gccggctgca gcgagaaaca
     7561 tctgctggac aagcccttct acaacgactt cgagcgggga gccgtggact cctacgatgt
     7621 gaccgtggac gaggaactgg gcgagatcca gctggtccgc atcgagaagc ggaagtactg
     7681 gctgaacgac gactggtatc tgaagtacat caccctgaaa accccccacg gcgactacat
     7741 cgagttccct tgctaccggt ggattacagg agacgtggaa gtggtgctgc gggacggcag
     7801 agccaagctg gccagggacg accagatcca catcctgaag cagcaccggc ggaaagagct
     7861 ggaaacccgg cagaaacagt accgctggat ggaatggaac cccggcttcc ccctgtccat
     7921 cgacgccaag tgccacaagg acctgccccg ggacatccag ttcgacagcg agaagggcgt
     7981 ggacttcgtg ctgaactaca gcaaggccat ggaaaacctg ttcatcaacc ggttcatgca
     8041 catgttccag agcagctgga acgacttcgc cgacttcgag aagatcttcg tgaagatcag
     8101 caacaccatc agcgagagag tgatgaacca ctggcaggaa gatctgatgt tcggctacca
     8161 gttcctgaac ggctgcaacc ccgtgctgat ccggcggtgt accgagctgc ctgagaagct
     8221 gcccgtgacc accgagatgg tggaatgcag cctggaacgg cagctgagcc tggaacagga
     8281 agtgcagcag ggcaacatct tcatcgtgga tttcgagctg ctggacggca ttgacgccaa
     8341 caagaccgac ccctgcaccc tgcagtttct ggccgctccc atctgcctgc tgtacaagaa
     8401 cctggccaac aagatcgtgc ctatcgccat ccagctgaac cagatccccg gcgacgagaa
     8461 ccccatcttc ctgcccagcg acgccaaata cgactggctg ctggccaaga tctgggtccg
     8521 ctccagcgac ttccacgtgc accagaccat cacccatctg ctgcggaccc acctggtgtc
     8581 cgaggtgttc ggaatcgcca tgtacagaca gctgcccgcc gtgcacccta tcttcaagct
     8641 gctggtggcc cacgtgcggt tcacaatcgc catcaacacc aaggccagag agcagctgat
     8701 ctgcgagtgc ggcctgttcg acaaggccaa cgccacaggc ggcggaggcc acgtgcagat
     8761 ggtgcagaga gccatgaagg acctgaccta cgccagcctg tgcttccccg aggccatcaa
     8821 ggccaggggc atggaaagca aagaggacat cccctactac ttctaccggg acgacggcct
     8881 gctggtctgg gaggccatca gaaccttcac cgccgaggtg gtggacatct actacgaggg
     8941 cgaccaggtg gtggaagagg accccgagct gcaggacttc gtgaacgacg tgtacgtgta
     9001 cggcatgcgg ggcagaaaga gcagcggctt ccccaagagc gtgaagtccc gcgagcagct
     9061 gagcgagtac ctgaccgtgg tgatcttcac cgccagcgcc cagcacgccg ccgtgaactt
     9121 cggccagtac gactggtgca gctggatccc caacgcccct cccaccatga gagccccccc
     9181 acctacagcc aagggcgtgg tgacaatcga gcagatcgtg gacaccctgc ccgaccgggg
     9241 cagaagctgt tggcatctgg gagccgtgtg ggccctgagc cagttccagg aaaacgagct
     9301 gttcctgggc atgtaccccg aggaacactt tatcgagaag cccgtgaaag aagccatggc
     9361 ccggttccgg aagaacctgg aagccatcgt gtccgtgatc gccgagcgga acaagaagaa
     9421 gcagctgccc tactactacc tgtcccccga cagaatccct aacagcgtgg ccatctgagc
     9481 ggcc
//
