LOCUS       sequence-101297-        7652 bp    DNA     circular SYN 15-APR-2025
DEFINITION  lentiviral expression of nuclear envelope transmembrane protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7652)
  TITLE     Schirmer lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NET29B.mm pRRLSIN (1744)
FEATURES             Location/Qualifiers
     source          1..7652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             236..416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1971..2481
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2400..2418
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    3506..4094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3559..3579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3977..3988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4181..4414
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4486..4607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4523..4542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4577..4596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4626..4761
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4793..4812)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4794..4812)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4822..4839)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4822..4838)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4831..4853)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4983..5438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5070..5089)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5280..5301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5467..5571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5572..5640,5641..6432)
                     /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(5790..5809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6603..7191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7092..7111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7345..7362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7479..7500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7515..7520,7521..7538,7539..7545)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7553..7569
                     /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     7558..7580
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7577..7593
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7577..7593
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7612..7632
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7614..7632
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     1921 ttttatcgat cacgagacta gcctcgagaa gcttgatatc gaattcccac ggggttgggg
     1981 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     2041 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     2101 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     2161 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     2221 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     2281 cgatgggctg tggccaatag cggctgctca gcggggcgcg ccgagagcag cggccgggaa
     2341 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     2401 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     2461 atcaccgacc tctctcccca ggggatgtcc ggccagctgg agcgttgcga gcgcgagtgg
     2521 cacgagctgg agggagaatt tcaggagctg caggagacac acaggatcta taagcagaag
     2581 ctggaggagc tgacctccct gcagacgctg tgcagcacct ccatcagcaa gcagaagagg
     2641 cacctgaagg acctgaagca cacactccag aggtacaagc gccattccag ccatgaggag
     2701 gcggcgctca tccaacagat gaccgccaac attaaggagc gacagaatgt cttctttgac
     2761 atggaggcct acctaccgaa gaagaatggg ctctacttga atctggttct tggcaacgtg
     2821 agcgtgaccc tcctcagcaa tcaggccaag tttgcttaca aggatgagta tgagaagttt
     2881 aagctctacc tgaccataat tcttcttctg ggtgctgtgg cgtgtcgatt tgtccttcat
     2941 tacagggtga ccgatgaagt cttcaacttc ctgctcgtgt ggtattactg caccttgacc
     3001 attcgagaga gcatcctcat cagcaatggt tccagaatca agggctggtg ggtgtctcac
     3061 cattatgtct ccacgttcct gtctggagtg atgctgactt ggcctaatgg actaatttac
     3121 cagaagtttc gaaatcagtt tttagcgttc tccatttttc agagctgcgt tcagtttcta
     3181 cagtactatt accagagggg ctgtctctac agactgcggg ccctggggga gaggaaccac
     3241 ctggacctca cagtggaagg gttccaatcc tggatgtggc ggggcctcac cttcctcctg
     3301 cctttcctct tctgtggcca tttctggcag ctctacaatg ccgtcaccct gtttgagctc
     3361 tccacccatg aggaatgcaa agaatggcag gtattcgtgc tggctctcac cttcctcatc
     3421 ctcttcctgg gcaattttct gaccacgctc aaagttgtcc atgccaaact ccacaagaac
     3481 agaaacaaga caaagcagcc atgacaatca acctctggat tacaaaattt gtgaaagatt
     3541 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3601 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3661 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3721 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3781 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3841 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3901 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3961 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4021 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4081 ggccgcctcc ccgcctggaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca
     4141 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     4201 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     4261 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     4321 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     4381 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     4441 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     4501 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4561 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     4621 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4681 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4741 gaggaggctt ttttggaggc ctaggctttt gcgtcgagac gtacccaatt cgccctatag
     4801 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4861 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4921 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4981 cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     5041 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5101 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5161 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5221 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5281 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5341 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5401 taacgcgaat tttaacaaaa tattaacgtt tacaatttcc caggtggcac ttttcgggga
     5461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6721 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     6781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7261 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7321 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7381 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7441 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7501 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7561 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     7621 cctcactaaa gggaacaaaa gctggagctg ca
//
