LOCUS       sequence-241438-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HES4 with normal stop codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  TITLE     Lentiviral expression plasmids for NOTCH effectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8508)
  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            "Unpublished"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: pLenti6.3_HES4
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1802..1919
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1942..2245
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2246..2449
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2400..2420
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2446..2470
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2566..2590
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2605..2614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3367..3391)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3444..3485
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3509..4097
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3562..3582)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3980..3991)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4112..4428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4279..4414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4476..4523
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4542..4940
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5027..5260
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5332..5453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5369..5388)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5423..5442
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5493..5628
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5648..5667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5649..5667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5677..5694)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5677..5693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5686..5708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5835..6290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5922..5941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6132..6153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6316..6420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6421..6489,6490..7281)
                     /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(6639..6658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7452..8040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7941..7960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8194..8211
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8328..8349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8364..8369,8370..8387,8388..8394)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8402..8418
                     /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     8407..8429
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8426..8442
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8426..8442
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8461..8481
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8463..8481
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta
     1921 tcgataagct tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     1981 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2041 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2101 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2161 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2221 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2281 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2341 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2401 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     2461 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact
     2521 ctagaggatc cactagtcca gtgtggtgga attctgcaga tatcaacaag tttgtacaaa
     2581 aaagcaggct ccgaattcgc ccttgccacc atggccgcag acacgccggg gaaaccgagc
     2641 gcctcgccga tggcaggagc gccggccagc gccagccgga ccccagacaa gccccggagc
     2701 gcggccgagc accgcaaggt ggggtcccgg ccgggcgtga ggggggcgac cggggggcgg
     2761 gagggacgcg ggactcagcc ggtgcccgac ccgcagtcct ccaagccggt catggagaag
     2821 cggcgccgag cgcgtattaa cgagagcctc gctcagctca aaaccctcat cctggacgcc
     2881 ctcagaaaag agagctcccg ccactcgaag ctggagaagg cggacatcct ggagatgacc
     2941 gtgagacacc tgcggagcct gcgtcgcgtg caggtgacgg ccgcgctcag cgccgacccc
     3001 gccgttctgg gcaagtaccg cgccggcttc cacgagtgtc tggcggaggt gaaccgcttc
     3061 ctggccggct gcgagggcgt cccggccgac gtgcgctccc gcctgctggg ccacctggca
     3121 gcctgcctgc gccagctggg accctcccgc cgcccggcct cgctgtcccc ggctgccccc
     3181 gcagaggccc cagcgcccga ggtctacgcg ggccgcccgc tgctgccatc gctcggcggc
     3241 cccttccctc tgctcgcgcc gccgctgctg ccgggtctga cccgggcgct gcccgccgcc
     3301 cccagggcgg ggccgcaggg cccgggtggg ccctggaggc cgtggctgcg ctgaagggcg
     3361 aattcgaccc agctttcttg tacaaagtgg ttgatatcca gcacagtggc ggccgctcga
     3421 gtctagaggg cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt
     3481 ctacgcgtac cggttagtaa tgatcgacaa tcaacctctg gattacaaaa tttgtgaaag
     3541 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     3601 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     3661 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     3721 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     3781 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     3841 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     3901 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3961 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4021 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     4081 ttgggccgcc tccccgcctg gcgatggtac cggtgtggaa agtccccagg ctccccagca
     4141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4261 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4321 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4381 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4441 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
     4501 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag cctttgtctc
     4561 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     4621 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     4681 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     4741 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     4801 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     4861 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     4921 gttatgtgtg ggagggctaa gcacaattcg agctcggtac ctttaagacc aatgacttac
     4981 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     5041 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     5101 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     5161 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     5221 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     5281 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     5341 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5401 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5461 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5521 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5581 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     5641 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     5701 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5761 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     5821 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5881 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5941 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6001 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6061 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6181 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6241 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     6301 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6361 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6421 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6481 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6541 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6601 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6661 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6721 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6781 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6841 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6901 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6961 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7021 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7081 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7141 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7201 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7261 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     8041 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8101 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     8161 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     8221 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     8281 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     8341 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     8401 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     8461 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaa
//
