LOCUS       sequence-148087-        8960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of genes (PGK 
            promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8960)
  AUTHORS   Morsut L, Roybal KT, Xiong X, Gordley RM, Coyle SM, Thomson M, Lim 
            WA
  TITLE     Engineering Customized Cell Sensing and Response Behaviors Using 
            Synthetic Notch Receptors.
  JOURNAL   Cell. 2016 Feb 11;164(4):780-91. doi: 10.1016/j.cell.2016.01.012. 
            Epub 2016 Jan 28.
  PUBMED    26830878
REFERENCE   2  (bases 1 to 8960)
  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"; doi: 
            "10.1016/j.cell.2016.01.012"; journalName: "Cell"; date: 
            "2016-02-11- 11"; volume: "164"; issue: "4"; pages: "780-91"
COMMENT     Sequence Label: pHR_PGK
FEATURES             Location/Qualifiers
     source          1..8960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             61..294
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(319..337)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(320..337)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(627..646)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     980..1002
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1040..1058)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1126..1230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1231..1299,1300..2091)
                     /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(1449..1468)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2262..2850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2751..2770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3004..3021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3091..3111)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3096..3425
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3276..3411
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4559..4624
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4754..4774
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5199..5333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5236..5255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5290..5309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5502..6135
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6182..6307
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6799..7032
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7217..7261
                     /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    7544..7661
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7721..8220
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7756..7779)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8153..8172
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     misc_feature    8370..8958
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8423..8443)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8841..8852)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 cgataccgtc gaccaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
       61 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
      121 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      181 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      241 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct
      301 agaattaatt ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt
      361 tatgtattaa ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc
      421 tggcttactg aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt
      481 tggacgaacc ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc
      541 aatcagcagg gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac
      601 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
      661 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
      721 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
      781 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
      841 tcgaatggtg cactctcagt acaatctagc tctgatgccg catagttaag ccagccccga
      901 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      961 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     1021 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1081 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1141 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1201 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1261 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1321 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1381 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1441 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1501 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1561 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1621 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1681 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1741 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1801 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1861 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1921 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1981 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2041 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2101 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2161 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2401 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2941 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3001 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3061 gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3241 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3301 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3361 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3421 gcaaaaagct tggacacaag acaggcttgc gagatatgtt tgagaatacc actttatccc
     3481 gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat gtgaaatact ggtttttagt
     3541 gcgccagatc tctataatct cgcgcaacct attttcccct cgaacacttt ttaagccgta
     3601 gataaacagg ctgggacact tcacatgagc gaaaaataca tcgtcacctg ggacatgttg
     3661 cagatccatg cacgtaaact cgcaagccga ctgatgcctt ctgaacaatg gaaaggcatt
     3721 attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact ggcgcgtgaa ctgggtattc
     3781 gtcatgtcga taccgtttgt atttccagct acgatcacga caaccagcgc gagcttaaag
     3841 tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt tattgatgac ctggtggata
     3901 ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa agcgcacttt gtcaccatct
     3961 tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt tgttgatatc ccgcaagata
     4021 cctggattga acagccgtgg gatatgggcg tcgtattcgt cccgccaatc tccggtcgct
     4081 aatcttttca acgcctggca ctgccgggcg ttgttctttt taacttcagg cgggttacaa
     4141 tagtttccag taagtattct ggaggctgca tccatgacac aggcaaacct gagcgaaacc
     4201 ctgttcaaac cccgctttaa acatcctgaa acctcgacgc tagtccgccg ctttaatcac
     4261 ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa ccatccctca ctggtatcgc
     4321 atgattaacc gtctgatgtg gatctggcgc ggcattgacc cacgcgaaat cctcgacgtc
     4381 caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg atgatttata cgatacggtg
     4441 attggctacc gtggcggcaa ctggatttat gagtgggccc cggatctttg tgaaggaacc
     4501 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
     4561 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
     4621 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga
     4681 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca
     4741 acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact ttccttcaga
     4801 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat
     4861 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt
     4921 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca
     4981 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt
     5041 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa
     5101 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     5161 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     5221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
     5341 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
     5401 attacctagt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     5461 agaaattgta tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc
     5521 ccaaagaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     5581 ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta
     5641 caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag
     5701 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg
     5761 gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt
     5821 caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag
     5881 gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc
     5941 tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6001 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6061 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     6121 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     6181 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     6241 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     6301 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     6361 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     6421 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     6481 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     6541 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     6601 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gtgatcttca
     6661 gacctggacg atatatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     6721 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     6781 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     6841 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     6901 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     6961 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     7021 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     7081 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     7141 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     7201 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     7261 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     7321 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     7381 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     7441 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     7501 gaacggatct cgacggtcgc caaatggcag tattcatcca caattttaaa agaaaagggg
     7561 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     7621 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     7681 gcagagatcc agtttggatc gataagcttg atatcgaatt gggtagggga ggcgcttttc
     7741 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     7801 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     7861 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     7921 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     7981 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     8041 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     8101 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     8161 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     8221 aattctcacg cgtcaagtgg agcaaggcag gtggacagtg gatccttgac ttgcggccgc
     8281 aactcccacc tgcaacatgc gtgactgact gaggccgcga ctctagagtc gacctgcagg
     8341 catgcaagct tgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     8401 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     8461 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     8521 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     8581 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     8641 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     8701 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     8761 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     8821 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     8881 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     8941 tttgggccgc ctccccgcat
//
