LOCUS       sequence-467818-        7143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty lentiviral backbone with C-terminal HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7143)
  AUTHORS   Addgene Research Program
  TITLE     Addgene Lentiviral Backbone Set
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7143)
  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            "Unpublished"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 C-HA Puro
FEATURES             Location/Qualifiers
     source          1..7143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..69
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        51..69
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        97..323
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             324..504
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    551..676
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1169..1402
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1587..1631
                     /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    1929..2046
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2461..2487
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2497..2553
                     /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             2554..3153
                     /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(2554..2573)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3050..3070
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3175..3763
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3228..3248)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3646..3657)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3843..4076
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4154..4275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4191..4210)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4245..4264
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4321..4456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4476..4495)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4477..4495)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4527..4544)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4527..4543)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4536..4558)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4752..4771)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4871..4893
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4931..4949)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5017..5121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5122..5190,5191..5982)
                     /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(5340..5359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6153..6741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6642..6661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6895..6912
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7029..7050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7065..7070,7071..7088,7089..7095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7103..7119
                     /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     7108..7130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7127..7143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7127..7143
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgattacg ccaagcttgc atgcctgcag catgattacg ccaagcgcgc aattaaccct
       61 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
      121 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
      181 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
      241 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
      301 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
      361 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
      421 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
      481 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
      541 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
      601 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
      661 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
      721 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
      781 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
      841 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
      901 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
      961 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     1021 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1081 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1141 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1201 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1261 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1321 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1381 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1441 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     1501 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     1561 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     1621 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     1681 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     1741 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     1801 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     1861 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     1921 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1981 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2041 aattttcggg tttattacag ggacagcaga gatccagttt ggttaattag ctagctaggt
     2101 cttgaaagga gtgggaattg gctccggtgc ccgtcagttt aattaagtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct tctagaggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc
     2401 ttcgaaggcg cgccgcgatc gcgatatcat gcattgatca ctcgagggcc ggccgggccc
     2461 tacccatacg atgttccgga ttacgctgga tccggcgcaa caaacttctc tctgctgaaa
     2521 caagccggag atgtcgaaga gaatcctgga ccgatgaccg agtacaagcc cacggtgcgc
     2581 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     2641 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     2701 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     2761 ggcgcagcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     2821 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     2881 gaaggcctcc tggcaccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     2941 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     3001 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     3061 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     3121 acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt aagtcgacat cgataatcaa
     3181 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3241 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3301 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3361 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3421 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3481 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3541 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     3601 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3661 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3721 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtttaaa cgtcgacttt
     3781 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     3841 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     3901 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     3961 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     4021 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     4081 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     4141 tgagaggtgt acaaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4201 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4261 catcaatgta tcttaactag ttcatgtctg gctctagcta tcccgcccct aactccgccc
     4321 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4381 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4441 ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc
     4501 gctccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
     4561 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     4621 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4681 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     4741 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4801 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4861 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4921 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4981 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5041 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5101 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5161 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5221 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5281 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5341 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5401 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5461 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5521 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5581 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5641 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5701 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5761 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5821 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5881 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5941 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6121 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6181 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6241 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6301 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6361 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6421 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6481 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6541 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6601 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6661 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6721 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6781 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6841 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6901 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6961 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7021 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7081 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7141 gac
//
