LOCUS       sequence-148715-        7547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of HA tagged Ubiquitin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7547)
  TITLE     Lentiviral expression
  JOURNAL   Unpublished
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: 
            "Unpublished"
COMMENT     Sequence Label: pLenti puro HA-Ubiquitin
FEATURES             Location/Qualifiers
     source          1..7547
                     /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"
     primer_bind     1929..1949
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        2238..2441
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2392..2412
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2489..2515
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2525..2752
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             2525..2752
                     /codon_start=1
                     /gene="Homo sapiens UBB"
                     /product="human ubiquitin B"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     misc_feature    2834..2951
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3000..3510
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3429..3447
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3532..4131
                     /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(3532..3551)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4028..4048
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4259..4492
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4564..4685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4601..4620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4655..4674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4725..4860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4880..4899)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4881..4899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4931..4948)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4931..4947)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4940..4962)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5156..5175)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5275..5297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5335..5353)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5421..5525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5526..5594,5595..6386)
                     /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(5744..5763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6557..7145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7046..7065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7299..7316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7433..7454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7469..7474,7475..7492,7493..7499)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7507..7523
                     /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     7512..7534
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7531..7547
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7531..7547
                     /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 gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc
     1921 ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     1981 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     2041 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     2101 actatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2161 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2221 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2281 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2341 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2401 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     2461 tccgctagcg ctaccggtcg ccaccatgta cccatacgat gttccggatt acgctagcgg
     2521 atccatgcag atcttcgtga aaacccttac cggcaagacc atcacccttg aggtggagcc
     2581 cagtgacacc atcgaaaatg tgaaggccaa gatccaggat aaggaaggca ttccccccga
     2641 ccagcagagg ctcatctttg caggcaagca gctggaagat ggccgtactc tttctgacta
     2701 caacatccag aaggagtcga ccctgcacct ggtcctgcgt ctgagaggtg gttattagcc
     2761 ggactcagat ctcgagctca agcttctcta gagaattctc gacctcgaga caaatggcag
     2821 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     2881 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     2941 ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggccg cggctcgagg
     3001 gggttggggt tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg cggctgctct
     3061 gggcgtggtt ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc
     3121 gttcgcagcg tcacccggat cttcgccgct acccttgtgg gccccccggc gacgcttcct
     3181 gctccgcccc taagtcggga aggttccttg cggttcgcgg cgtgccggac gtgacaaacg
     3241 gaagccgcac gtctcactag taccctcgca gacggacagc gccagggagc aatggcagcg
     3301 cgccgaccgc gatgggctgt ggccaatagc ggctgctcag cagggcgcgc cgagagcagc
     3361 ggccgggaag gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg
     3421 cccgcgcggt gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt cggctccctc
     3481 gttgaccgaa tcaccgacct ctctccccag ggggatccac cggagcttac catgaccgag
     3541 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3601 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3661 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3721 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3781 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3841 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     3901 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3961 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4021 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4081 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     4141 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcggt acctttaaga
     4201 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     4261 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     4321 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     4381 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     4441 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     4501 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     4561 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4621 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4681 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     4741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4801 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4861 tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctccc cgggtaccga
     4921 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4981 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5041 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     5101 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     5161 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5221 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5281 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5341 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7261 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7321 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7381 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7441 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7501 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgac
//
