LOCUS       sequence-149803-        8410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EglN3-H196A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8410)
  TITLE     Schlisio lab plasmids
REFERENCE   2  (bases 1 to 8410)
  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-H196A
FEATURES             Location/Qualifiers
     source          1..8410
                     /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..4001
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3466..3486)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3884..3895)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4016..4332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4183..4318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4380..4427
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4446..4844
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4931..5164
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5236..5357
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5273..5292)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5327..5346
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5397..5532
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5552..5571)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5553..5571)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5581..5598)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5581..5597)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5590..5612)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5739..6194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5826..5845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6036..6057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6220..6324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6325..6393,6394..7185)
                     /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(6543..6562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7356..7944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7845..7864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8098..8115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8232..8253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8268..8273,8274..8291,8292..8298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8306..8322
                     /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     8311..8333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8330..8346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8330..8346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8365..8385
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8367..8385
                     /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 gaacccagcc 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 cctctggatt acaaaattgg aaagattgac tggtattctt aactatgttg ctccttttac
     3481 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3541 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3601 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3661 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3721 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3781 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3841 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3901 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3961 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggcgatg gtaccggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4141 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4201 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4261 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4321 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     4381 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     4441 aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     4501 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     4561 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     4621 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     4681 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     4741 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     4801 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcacaa ttcgagctcg
     4861 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     4921 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     4981 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5041 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5101 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5161 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     5221 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5281 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5341 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     5401 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5461 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5521 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     5581 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5641 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5701 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     5761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5881 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5941 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6061 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6181 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6241 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6301 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6361 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6421 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6481 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6541 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6601 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6661 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6721 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6781 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6841 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6901 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6961 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7021 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7081 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7141 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7201 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7261 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7321 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7381 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7441 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7501 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7561 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7621 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7681 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7741 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7801 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7861 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7921 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7981 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8041 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8101 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8161 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8221 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8281 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8341 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
     8401 ctgcaagctt
//
