LOCUS       sequence-416829-        8405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  doxycycline-inducible expression of Human inserts (MCS) in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8405)
  AUTHORS   Henriques-Oliveira L, Altman AR, Kurochkin I, Ascic E, Halitzki E, 
            Matei A, Pertiga-Cabral D, Ulmert I, Holst S, Nair MS, Cunha PP, 
            Park S, Vergani S, Kharas MG, Yuan J, Lahl K, Rosa FF, Pires CF, 
            Pereira C-F
  TITLE     Anchored screening identifies transcription factor blueprints 
            underlying dendritic cell diversity and subset-specific anti-tumor 
            immunity
  JOURNAL   Immunity. 58, 1-20. October 2025.
REFERENCE   2  (bases 1 to 8405)
  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: "Immunity. 
            58, 1-20. October 2025."
COMMENT     Sequence Label: pFUW-TetO-MCS
FEATURES             Location/Qualifiers
     source          1..8405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        408..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             624..804
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    851..976
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1469..1702
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1887..1931
                     /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    2229..2346
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2403..2421,2422..2444,2445..2463,2464..2486,2487..2505,
                     2506..2528,2529..2547,2548..2570,2571..2589,2590..2612,
                     2613..2631,2632..2654,2655..2673)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2706..2744
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2741..2765
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    2886..3474
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2939..2959)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3357..3368)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3477..3493)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3478..3494)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3999..4179
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4205..4222)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4211..4435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4481..4909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4568..4587)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4778..4799
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4918..4938)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4923..5252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5103..5238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5165..5184
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5300..5347
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5366..5740
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5870..5991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5907..5926)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5961..5980
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6040..6056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6040..6056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6053..6075)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6064..6080
                     /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)."
     promoter        complement(join(6088..6094,6095..6112,6113..6118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6133..6154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6271..6288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6442..7030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6522..6541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7201..7992,7993..8061))
                     /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     7824..7843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8062..8166)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8241..8260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
      661 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
      721 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
      781 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
      841 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
      901 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
      961 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     1021 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     1081 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     1141 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     1201 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     1261 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     1321 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1381 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1441 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1501 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1561 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1621 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1681 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1741 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     1801 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     1861 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     1921 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     1981 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     2041 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     2101 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     2161 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     2221 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2281 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2341 aattttcggg tttattacag ggacagcaga gatccagttt ggttagatct cgagtttacc
     2401 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     2461 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     2521 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     2581 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     2641 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagct cggtacccgg
     2701 gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc
     2761 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     2821 ccagcctccg cggccccgaa ttcaccggtt ctagagctag cttcgaagat atcaagctta
     2881 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2941 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3001 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3061 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3121 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3181 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3241 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3301 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3361 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3421 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3481 accgtcgacc tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     3541 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     3601 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     3661 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     3721 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     3781 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     3841 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     3901 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     3961 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
     4021 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     4081 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     4141 ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga
     4201 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4261 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4321 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4381 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4441 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4501 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4561 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4621 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4681 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4741 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4801 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4861 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4921 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4981 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5041 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5101 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5161 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5221 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5281 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     5341 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     5401 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     5461 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
     5521 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
     5581 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
     5641 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
     5701 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
     5761 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5821 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5881 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5941 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6001 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6061 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6121 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6181 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6241 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6301 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6361 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6421 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6481 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6541 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6601 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6661 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6721 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6781 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6841 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6901 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7021 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7081 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7141 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7201 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7261 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7321 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7381 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7441 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7501 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7561 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7621 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7681 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7741 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7801 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7861 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7921 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7981 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8041 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8101 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8161 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     8221 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     8281 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8341 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     8401 cttcg
//
