LOCUS       sequence-348944-        7547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of anti-HIV-1 MT-C34 peptide on 
            cell surface.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7547)
  AUTHORS   Maslennikova A, Kruglova N, Kalinichenko S, Komkov D, Shepelev M, 
            Golubev D, Siniavin A, Vzorov A, Filatov A, Mazurov D
  TITLE     Engineering T-Cell Resistance to HIV-1 Infection via Knock-In of 
            Peptides from the Heptad Repeat 2 Domain of gp41.
  JOURNAL   mBio. 2022 Jan 25:e0358921. doi: 10.1128/mbio.03589-21.
  PUBMED    35073736
REFERENCE   2  (bases 1 to 7547)
  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: "mBio. 2022 
            Jan 25:e0358921. doi: 10.1128/mbio.03589-21."
COMMENT     Sequence Label: pUCHR-mClover-MT-C34
FEATURES             Location/Qualifiers
     source          1..7547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    57..182
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    675..908
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1093..1137
                     /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    1432..1549
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1609..1912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1913..2116
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2067..2087
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2113..2137
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2222..2224,2225..2227,2228..2938)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2267..2288)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2875..2896
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3074..3130
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     LTR             3465..4098
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4147..4163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4147..4163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4160..4182)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4171..4187
                     /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)."
     promoter        complement(join(4195..4201,4202..4219,4220..4225))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4240..4261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4378..4395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      4538..4673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(4736..5324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4816..4835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5495..6286,6287..6355))
                     /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     6118..6137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6356..6460)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6528..6546
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6584..6606)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6706..6725
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6919..6941
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6933..6950
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6934..6950
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             join(7300..7547,1..10)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
       61 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
      121 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
      181 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
      241 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
      301 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
      361 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
      421 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
      481 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
      541 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
      601 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
      661 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
      721 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
      781 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
      841 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
      901 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
      961 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     1021 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     1081 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     1141 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     1201 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     1261 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     1321 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     1381 ggatctcgac ggtatcggac cctcgagaca aatggcagta ttcatccaca attttaaaag
     1441 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     1501 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     1561 cagggacagc agaaatccac tttgggtccg ataagcttgg gagttccgcg ttacataact
     1621 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     1681 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1741 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     1801 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1861 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1921 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1981 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2041 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2101 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     2161 ttttgacctc catagaagac accgggaccg atccagcctc cgaattcgca ggtcgaccac
     2221 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2281 cggcgacgta aacggccaca agttcagcgt ccgcggcgag ggcgagggcg atgccaccaa
     2341 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2401 cctcgtgacc accttcggct acggcgtggc ctgcttcagc cgctaccccg accacatgaa
     2461 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctc
     2521 tttcaaggac gacggtacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2581 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2641 caagctggag tacaacttca acagccacaa cgtctatatc acggccgaca agcagaagaa
     2701 cggcatcaag gctaacttca agatccgcca caacgttgag gacggcagcg tgcagctcgc
     2761 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2821 ctacctgagc catcagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     2881 cctgctggag ttcgtgaccg ccgccgggat tacacatggc atggacgagc tgtacaagcc
     2941 agccaaacct ccggccaagg cacaagttgt gggatggcca ccggtgagat cataccggaa
     3001 gaacgtgatg gtttcctgcc aaaaatcaag cggtggcccg gaggcggcgg cgttcgtgaa
     3061 agtaggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa
     3121 tcctggaccg aagcgcttcc tcttcctcct actcaccatc agcctcctgg ttatggtaca
     3181 gatacaaact ggactctcaa tgacctggat ggagtgggac cgggagatca ataactacac
     3241 ctcactgatc cacagcctga tcgaggagtc ccagaaccag caggagaaga acgagcagga
     3301 gctcctgtca gcatccagca acataagcgg aggcattttc cttttcttcg tggccaatgc
     3361 cataatccac ctcttctgct tcagttgagc tcgagtacct ttaagaccaa tgacttacaa
     3421 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     3481 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     3541 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
     3601 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
     3661 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
     3721 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggagta
     3781 cttcaagaac tgctgacatc gagcttgcta caagggactt tccgctgggg actttccagg
     3841 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatgctgca tataagcagc
     3901 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     3961 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     4021 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     4081 tgtggaaaat ctctagcagg atcctctaga gtcgacctgc aggcatgcaa gcttggcgta
     4141 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4201 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4261 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4321 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4381 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4441 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgcatctcaa
     4501 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4561 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4621 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagatgt
     4681 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4741 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4801 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4861 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4921 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4981 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5041 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5101 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5161 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5221 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5281 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5341 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5401 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5461 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     5521 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     5581 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5641 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5701 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5761 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5821 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5881 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5941 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6001 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6061 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6121 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6181 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6241 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6301 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6361 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6421 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6481 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     6541 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6601 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6661 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     6721 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     6781 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6841 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     6901 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgatat
     6961 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
     7021 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     7081 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
     7141 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
     7201 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
     7261 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
     7321 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
     7381 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7441 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7501 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaa
//
