LOCUS       sequence-467817-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty lentiviral backbone with N-terminal HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  AUTHORS   Addgene Research Program
  TITLE     Addgene Lentiviral Backbone Set
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7146)
  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            "Unpublished"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: pAG Lenti CMV N-HA Puro
FEATURES             Location/Qualifiers
     source          1..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(138..157)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     257..279
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(317..335)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        403..507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(508..576,577..1368)
                     /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(726..745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1539..2127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2028..2047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2281..2298
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2415..2436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2451..2456,2457..2474,2475..2481)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2489..2505
                     /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     2494..2516
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2513..2529
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2513..2529
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2578..2598
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2580..2598
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2626..2852
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2853..3033
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3080..3205
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3698..3931
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4116..4160
                     /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    4458..4575
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4676..4879
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4830..4850
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4933..4959
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5029..5085
                     /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"
     CDS             5086..5685
                     /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(5086..5105)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5582..5602
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5707..6295
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5760..5780)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6178..6189)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6375..6608
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6686..6807
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6723..6742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6777..6796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6853..6988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7008..7027)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7009..7027)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7059..7076)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7059..7075)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7068..7090)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
       61 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
      121 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
      181 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
      241 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
      301 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
      361 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      421 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
      481 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
      541 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
      601 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
      661 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
      721 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
      781 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
      841 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
      901 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
      961 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1021 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1081 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1141 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1201 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1261 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1321 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     1381 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2221 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2281 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     2341 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2401 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     2461 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     2521 agctatgacc atgattacgc caagcttgca tgcctgcagc atgattacgc caagcgcgca
     2581 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     2641 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     2701 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     2761 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     2821 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     2881 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     2941 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     3001 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     3061 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     3121 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     3181 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     3241 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     3301 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     3361 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     3421 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     3481 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     3541 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     3601 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     3661 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     3721 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     3781 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     3841 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     3901 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     3961 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     4021 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     4081 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     4141 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     4201 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     4261 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     4321 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     4381 acagatccat tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg
     4441 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     4501 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     4561 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gttaattagc
     4621 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagttta attaagtgat
     4681 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     4741 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     4801 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     4861 ggtctatata agcagagctt ctagaggttt agtgaaccgt cagatccgct agcgctaccg
     4921 gtcgccacca tgtacccata cgatgttccg gattatgctt tcgaaggcgc gccgcgatcg
     4981 cgggcccgat atctgatcac tcgagggccg gccatgcatg gatccggcgc aacaaacttc
     5041 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgatgac cgagtacaag
     5101 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     5161 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     5221 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     5281 gtcgcggacg acggcgcagc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     5341 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     5401 cagcaacaga tggaaggcct cctggcaccg caccggccca aggagcccgc gtggttcctg
     5461 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     5521 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     5581 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     5641 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac
     5701 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5761 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5821 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5881 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5941 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     6001 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     6061 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     6121 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     6181 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6241 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgttta
     6301 aacgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6361 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     6421 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6481 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6541 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6601 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6661 aatatcagag agtgagaggt gtacaaactt gtttattgca gcttataatg gttacaaata
     6721 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6781 tttgtccaaa ctcatcaatg tatcttaact agttcatgtc tggctctagc tatcccgccc
     6841 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6901 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6961 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     7021 cgtattacgc gcgctccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt
     7081 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     7141 gccagc
//
