LOCUS       sequence-241458-        8443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HES7 with normal stop codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8443)
  TITLE     Lentiviral expression plasmids for NOTCH effectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8443)
  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            "Unpublished"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_HES7
FEATURES             Location/Qualifiers
     source          1..8443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1944..2247
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2248..2451
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2402..2422
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2448..2472
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2568..2592
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2607..2616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3304..3328)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3381..3422
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3446..4034
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3499..3519)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3917..3928)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4049..4365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4216..4351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4413..4460
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4479..4877
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4964..5197
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5269..5390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5306..5325)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5360..5379
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5430..5565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5585..5604)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5586..5604)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5614..5631)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5614..5630)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5623..5645)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5772..6227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5859..5878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6069..6090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6253..6357
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6358..6426,6427..7218)
                     /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(6576..6595)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7389..7977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7878..7897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8131..8148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8265..8286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8301..8306,8307..8324,8325..8331)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8339..8355
                     /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     8344..8366
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8363..8379
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8363..8379
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8398..8418
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8400..8418
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1981 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2041 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2101 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2161 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2221 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2281 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     2341 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     2401 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
     2461 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga
     2521 ctctagagga tccactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca
     2581 aaaaagcagg ctccgaattc gcccttgcca ccatggtcac ccgggatcga gctgagaata
     2641 gggacggccc caagatgctc aagccgcttg tggagaagcg gcgccgggac cgcatcaacc
     2701 gcagcctgga agagctgagg ctgctgctgc tggagcggac ccgggaccag aacctccgga
     2761 acccgaagct ggagaaagcg gagatattgg agttcgccgt gggctacttg agggagcgaa
     2821 gccgggtgga gcccccgggg gttccccggt ccccagtcca ggacgccgag gcgctcgcca
     2881 gctgctactt gtccggtttc cgcgagtgcc tgcttcgctt ggcggccttc gcgcacgacg
     2941 ccagcccggc cgcccgcgcc cagctcttct ccgcgctgca cggctatctg cgccccaaac
     3001 cgccccggcc caagccggta gatccgaggc ctccagcgcc gcgcccatcc ctggaccccg
     3061 ccgcaccggc ccttggccct gcgctgcacc agcgcccccc agtgcaccag ggccacccta
     3121 gcccgcgctg cgcatggtcc ccatccctct gctccccgcg cgccggggat tctggcgcgc
     3181 cggcgcccct caccggactg ctgccgccgc caccgccgcc tcacagacaa gacggggcgc
     3241 ccaaggcccc gctgcccccg ccgcccgctt tctggagacc ttggccctga aagggcgaat
     3301 tcgacccagc tttcttgtac aaagtggttg atatccagca cagtggcggc cgctcgagtc
     3361 tagagggccc gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     3421 cgcgtaccgg ttagtaatga tcgacaatca acctctggat tacaaaattt gtgaaagatt
     3481 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3541 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3601 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3661 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3721 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3781 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3841 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3901 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3961 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4021 ggccgcctcc ccgcctggcg atggtaccgg tgtggaaagt ccccaggctc cccagcaggc
     4081 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4201 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4261 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4321 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4381 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     4441 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag
     4501 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     4561 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     4621 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     4681 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     4741 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     4801 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     4861 atgtgtggga gggctaagca caattcgagc tcggtacctt taagaccaat gacttacaag
     4921 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     4981 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5041 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5101 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5161 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     5221 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     5281 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5341 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5401 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5461 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5521 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     5581 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     5641 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     5701 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5761 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5821 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5881 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5941 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6001 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6061 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6121 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6181 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     6241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6301 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6361 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6421 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6481 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6541 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6601 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6661 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6721 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6781 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6841 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6901 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6961 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7021 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7081 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7141 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7201 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7261 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7321 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7381 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7441 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7501 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7561 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7621 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7681 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7741 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7801 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7861 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7921 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7981 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     8041 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     8101 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     8161 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     8221 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     8281 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     8341 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     8401 attaaccctc actaaaggga acaaaagctg gagctgcaag ctt
//
