LOCUS       sequence-449976-        7694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing human H2BC21 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7694)
  TITLE     Lentiviral plasmids expressing popular genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7694)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLenti puro HA-H2BC21
FEATURES             Location/Qualifiers
     source          1..7694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             108..288
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(289..515)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(543..563)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(543..561)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(612..628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(612..628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(625..647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    636..652
                     /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(660..666,667..684,685..690))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    705..726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(843..860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1014..1602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1094..1113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1773..2564,2565..2633))
                     /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     2396..2415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2634..2738)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2806..2824
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2862..2884)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2984..3003
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3197..3219
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3211..3228
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3212..3228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3260..3279
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3260..3278
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3299..3434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3474..3595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3485..3504)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3539..3558
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3667..3900
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(4028..4627)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4111..4131)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4608..4627
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4649..5159)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4712..4730)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    5208..5325
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5791..5817)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(5865..6068)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5894..5914)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(6357..6377)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(6675..6719)
                     /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    6904..7137
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    join(7630..7694,1..61)
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga
       61 gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt
      121 ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc
      181 acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag
      241 ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccgt ttatgtatcg
      301 agctaggcac ttaaatacaa tatctctgca atgcggcaat tcagtggttc gtccaatcca
      361 tgtcagaccc gtctgttgcc ttcctaataa ggcacgatcg taccacctta cttccaccaa
      421 tcggcatgca cggtgctttt tctctccttg taaggcatgt tgctaactca tcgttaccat
      481 gttgcaagac tacaagagta ttgcataaga ctacattaag cttgcagctc cagcttttgt
      541 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat gctgcaggca tgcaagcttg
      601 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      661 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      721 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      781 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      841 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      901 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      961 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1021 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1081 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1141 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1201 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1261 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1321 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1381 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1441 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1501 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1561 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1621 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1681 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1741 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1801 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1861 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1921 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1981 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2041 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2101 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2161 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2221 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2281 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2341 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2401 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2461 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2521 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2581 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2641 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2701 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2761 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2821 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2881 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2941 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     3001 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3061 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     3121 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     3181 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
     3241 ggtacccggg gagcgcgcgt aatacgactc actatagggc gaattgggta cgtccctagg
     3301 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     3361 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     3421 gttaggggcg ggatgggcgg agttaggggc gggatagcta gagccagaca tgataagata
     3481 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3541 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttcctct
     3601 cactctctga tattcatttc tttgcaagtt ataaatactg aataataaga tgacatgaac
     3661 tactactgct agagattttc cacactgact aaaagggtct gagggatctc tagttaccag
     3721 agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct
     3781 taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga
     3841 gagacccagt acaagcaaaa agcagatctt gtcttcgttg ggagtgaatt agcccttcca
     3901 gtcccccctt ttcttttaaa aagtggctaa gatctacagc tgccttgtaa gtcattggtc
     3961 ttaaaggtac cgatgcatgg ggtcgtgcgc tcctttcggt cgggcgctgc gggtcgtggg
     4021 gcgggcgtca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct
     4081 cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg
     4141 aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga
     4201 cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc
     4261 acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca
     4321 gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt
     4381 cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct
     4441 tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga
     4501 tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga
     4561 gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact
     4621 cggtcatggt aagctccggt ggatccccct ggggagagag gtcggtgatt cggtcaacga
     4681 gggagccgac tgccgacgtg cgctccggag gcttgcagaa tgcggaacac cgcgcgggca
     4741 ggaacagggc ccacactacc gccccacacc ccgcctcccg caccgcccct tcccggccgc
     4801 tgctctcggc gcgccctgct gagcagccgc tattggccac agcccatcgc ggtcggcgcg
     4861 ctgccattgc tccctggcgc tgtccgtctg cgagggtact agtgagacgt gcggcttccg
     4921 tttgtcacgt ccggcacgcc gcgaaccgca aggaaccttc ccgacttagg ggcggagcag
     4981 gaagcgtcgc cggggggccc acaagggtag cggcgaagat ccgggtgacg ctgcgaacgg
     5041 acgtgaagaa tgtgcgagac ccagggtcgg cgccgctgcg tttcccggaa ccacgcccag
     5101 agcagccgcg tccctgcgca aacccagggc tgccttggaa aaggcgcaac cccaaccccc
     5161 tcgagccgcg gccaaagtgg atctctgctg tccctgtaat aaacccgaaa attttgaatt
     5221 tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt
     5281 ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg gatgaatact
     5341 gccatttgtc tcgaggtcga gaattctcta gagaagcttg agctcgagat ctgagtccgg
     5401 ctatttgctt gaggtgtact tcgttacggc ctttgttccc tcagataccg cgtgtttcgc
     5461 cagctcccct ggaagaagaa gccggactgc agtttggatt tcccggcttg ttatggtact
     5521 tcgtttgtta taatgagcga gcctgcttgc ctcgccagcg attctttcaa aaatgtcgtt
     5581 tacaaatgag ttcattatcc ccattgcttt agaggagatg cccgtgtccg ggtgcacctg
     5641 tttcaagact ttataaacat atatagagta actttccttc ctgcttcgct tcctcttttt
     5701 tccgtctttt ttttgagctt tcgtaactgc ttttttagat cccttttttg gtgctggggc
     5761 tgatttcgct ggctcgggca tggatccgct agcgtaatcc ggaacatcgt atgggtacat
     5821 ggtggcgacc ggtagcgcta gcggatctga cggttcacta aaccagctct gcttatatag
     5881 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     5941 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     6001 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     6061 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     6121 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     6181 gggggcgtac ttggcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac
     6241 ccaagaaatt attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc
     6301 aaattaattc caattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc
     6361 atgggaaata ggccctcggt gaagggggcg gccgctcgag gctagtctcg tgatcgatac
     6421 cgtcgagatc cgttcactaa tcgaatggat ctgtctctgt ctctctctcc accttcttct
     6481 tctattcctt cgggcctgtc gggtcccctc ggggttggga ggtgggtctg aaacgataat
     6541 ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac tattcttaaa
     6601 cctaccaagc ctcctactat cattatgaat aattttatat accacagcca atttgttatg
     6661 ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg ttcattcttt
     6721 tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta attgttaatt
     6781 tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga tttattactc
     6841 caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca accccaaatc
     6901 cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc ctggagctgc
     6961 ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc cctcagcaaa
     7021 ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt cagcgtcatt
     7081 gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa agctcctatt
     7141 cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg tgctactcct
     7201 aatggttcaa tttttactac tttatattta tataattcac ttctccaatt gtccctcata
     7261 tctcctcctc caggtctgaa gatcagcggc cgcttgctgt gcggtggtct tacttttgtt
     7321 ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg
     7381 cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg
     7441 aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc
     7501 aggattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata
     7561 ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg
     7621 cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt
     7681 tggcgtactc acca
//
