LOCUS       sequence-496725-        5839 bp    DNA     circular SYN 08-JAN-2026
DEFINITION  Lentiviral vector for Gal4 inducible humanIL-10 with mCherry 
            reporter and constitutive tBFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5839)
  AUTHORS   Roybal KT, Williams JZ, Morsut L, Rupp LJ, Kolinko I, Choe JH, 
            Walker WJ, McNally KA, Lim WA
  TITLE     Engineering T Cells with Customized Therapeutic Response Programs 
            Using Synthetic Notch Receptors.
  JOURNAL   Cell. 2016 Oct 6;167(2):419-432.e16. doi: 
            10.1016/j.cell.2016.09.011. Epub 2016 Sep 29.
  PUBMED    27693353
REFERENCE   2  (bases 1 to 5839)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 8, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2016.09.011"; journalName: "Cell"; date: "2016-10-6-
            6"; volume: "167"; issue: "2"; pages: "419-432.e16"
COMMENT     Sequence Label: pHR_Gal4UAS_humanIL-10_IRES_mCherry_PGK_tBFP
FEATURES             Location/Qualifiers
     source          1..5839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(204..222)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(205..222)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(512..531)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(745..764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     864..886
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(924..942)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1010..1114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1115..1183,1184..1975)
                     /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(1333..1352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2146..2734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2635..2654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2888..2905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2975..2995)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2980..3309
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3160..3295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4443..4508
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4638..4658
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5083..5217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5120..5139)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5174..5193
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(5385..5839,1..179)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
       61 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      121 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      181 catctagaat taattccgtg tattctatag tgtcacctaa atcgtatgtg tatgatacat
      241 aaggttatgt attaattgta gccgcgttct aacgacaata tgtacaagcc taattgtgta
      301 gcatctggct tactgaagca gaccctatca tctctctcgt aaactgccgt cagagtcggt
      361 ttggttggac gaaccttctg agtttctggt aacgccgtcc cgcacccgga aatggtcagc
      421 gaaccaatca gcagggtcat cgctagccag atcctctacg ccggacgcat cgtggccggc
      481 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
      541 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
      601 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
      661 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
      721 gagcgtcgaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
      781 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      841 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
      901 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
      961 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1021 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1201 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1261 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1321 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1381 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1441 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1501 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1561 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1621 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1681 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1741 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1801 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1861 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1921 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1981 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2041 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2101 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2161 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2221 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2281 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2341 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2401 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2461 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2521 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2581 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2641 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2701 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2761 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2821 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2881 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2941 ccccgcgcgt tggccgattc attaatgcag ctgtggaatg tgtgtcagtt agggtgtgga
     3001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3061 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3121 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3181 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3241 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3301 ttttgcaaaa agcttggaca caagacaggc ttgcgagata tgtttgagaa taccacttta
     3361 tcccgcgtca gggagaggca gtgcgtaaaa agacgcggac tcatgtgaaa tactggtttt
     3421 tagtgcgcca gatctctata atctcgcgca acctattttc ccctcgaaca ctttttaagc
     3481 cgtagataaa caggctggga cacttcacat gagcgaaaaa tacatcgtca cctgggacat
     3541 gttgcagatc catgcacgta aactcgcaag ccgactgatg ccttctgaac aatggaaagg
     3601 cattattgcc gtaagccgtg gcggtctgta ccgggtgcgt tactggcgcg tgaactgggt
     3661 attcgtcatg tcgataccgt ttgtatttcc agctacgatc acgacaacca gcgcgagctt
     3721 aaagtgctga aacgcgcaga aggcgatggc gaaggcttca tcgttattga tgacctggtg
     3781 gataccggtg gtactgcggt tgcgattcgt gaaatgtatc caaaagcgca ctttgtcacc
     3841 atcttcgcaa aaccggctgg tcgtccgctg gttgatgact atgttgttga tatcccgcaa
     3901 gatacctgga ttgaacagcc gtgggatatg ggcgtcgtat tcgtcccgcc aatctccggt
     3961 cgctaatctt ttcaacgcct ggcactgccg ggcgttgttc tttttaactt caggcgggtt
     4021 acaatagttt ccagtaagta ttctggaggc tgcatccatg acacaggcaa acctgagcga
     4081 aaccctgttc aaaccccgct ttaaacatcc tgaaacctcg acgctagtcc gccgctttaa
     4141 tcacggcgca caaccgcctg tgcagtcggc ccttgatggt aaaaccatcc ctcactggta
     4201 tcgcatgatt aaccgtctga tgtggatctg gcgcggcatt gacccacgcg aaatcctcga
     4261 cgtccaggca cgtattgtga tgagcgatgc cgaacgtacc gacgatgatt tatacgatac
     4321 ggtgattggc taccgtggcg gcaactggat ttatgagtgg gccccggatc tttgtgaagg
     4381 aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta
     4441 aggtaaatat aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     4501 tattttagat tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg
     4561 aggaaaacct gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact
     4621 ctcaacattc tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt
     4681 cagaattgct aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg
     4741 ctatttacac cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt
     4801 ctgtaacctt tataagtagg cataacagtt ataatcataa catactgttt tttcttactc
     4861 cacacaggca tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct
     4921 ttttaatttg taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc
     4981 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     5041 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     5101 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5161 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
     5221 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
     5281 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
     5341 taaagaaatt gtatttgtta aatatgtact acaaacttag tagttggaag ggctaattca
     5401 ctcccaaaga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     5461 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     5521 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     5581 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     5641 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     5701 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     5761 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     5821 tgctttttgc ctgtactgg
//
