LOCUS       sequence-149799-        8409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EglN3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8409)
  TITLE     Schlisio lab plasmids
REFERENCE   2  (bases 1 to 8409)
  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"
COMMENT     Sequence Label: pLp6.3-EglN3
FEATURES             Location/Qualifiers
     source          1..8409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1944..2247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2248..2451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2402..2422
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2448..2472
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3350..3391
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3415..4000
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3465..3485)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3883..3894)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4015..4331
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4182..4317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4379..4426
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4445..4843
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4930..5163
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5235..5356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5272..5291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5326..5345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5396..5531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5551..5570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5552..5570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5580..5597)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5580..5596)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5589..5611)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5738..6193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5825..5844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6035..6056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6219..6323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6324..6392,6393..7184)
                     /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(6542..6561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7355..7943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7844..7863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8097..8114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8231..8252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8267..8272,8273..8290,8291..8297)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8305..8321
                     /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     8310..8332
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8329..8345
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8329..8345
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8364..8384
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8366..8384
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1981 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2041 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2101 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2161 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2221 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2281 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2341 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2401 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     2461 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga
     2521 ctctagagga tccactagtc cagtgtggtg gaattggccc ttatgcccct gggacacatc
     2581 atgaggctgg acctggagaa aattgccctg gagtacatcg tgccctgtct gcacgaggtg
     2641 ggcttctgct acctggacaa cttcctgggc gaggtggtgg gcgactgcgt cctggagcgc
     2701 gtcaagcagc tgcactgcac cggggccctg cgggacggcc agctggcggg gccgcgcgcc
     2761 ggcgtctcca agcgacacct gcggggcgac cagatcacgt ggatcggggg caacgaggag
     2821 ggctgcgagg ccatcagctt cctcctgtcc ctcatcgaca ggctggtcct ctactgcggg
     2881 agccggctgg gcaaatacta cgtcaaggag aggtctaagg caatggtggc ttgctatccg
     2941 ggaaatggaa caggttatgt tcgccacgtg gacaacccca acggtgatgg tcgctgcatc
     3001 acctgcatct actatctgaa caagaattgg gatgccaagc tacatggtgg gatcctgcgg
     3061 atatttccag aggggaaatc attcatagca gatgtggagc ccatttttga cagactcctg
     3121 ttcttctggt cagatcgtag gaacccacac gaagtgcagc cctcttacgc aaccagatat
     3181 gctatgactg tctggtactt tgatgctgaa gaaagggcag aagccaaaaa gaaattcagg
     3241 aatttaacta ggaaaactga atctgccctc actgaagact gaaagggcca attctgcaga
     3301 tatccagcac agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat
     3361 ccctaaccct ctcctcggtc tcgattctac gcgtaccggt tagtaatgat cgacaatcaa
     3421 cctctggata caaaattgga aagattgact ggtattctta actatgttgc tccttttacg
     3481 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3541 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3601 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3661 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3721 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3781 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     3841 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3901 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3961 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggcgatgg taccggtgtg
     4021 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4081 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4261 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4321 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     4381 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     4441 aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     4501 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     4561 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
     4621 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     4681 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
     4741 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     4801 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacaat tcgagctcgg
     4861 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     4921 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     4981 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5041 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5101 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5161 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     5221 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5281 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5341 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     5401 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5461 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5521 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     5581 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     5641 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     5701 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     5761 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5821 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5881 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5941 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6001 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     6061 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     6121 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     6181 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6241 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6301 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6361 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6421 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6481 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6541 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6601 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6661 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6721 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6781 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6841 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6901 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6961 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7021 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7081 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7141 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7201 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7261 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7321 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7381 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7441 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7501 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7561 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7621 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7681 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7741 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7801 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7861 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7921 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7981 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8041 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8101 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     8161 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     8221 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     8281 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     8341 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     8401 tgcaagctt
//
