LOCUS       sequence-244596-        8287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HES2 with normal stop codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8287)
  TITLE     Lentiviral expression plasmids for NOTCH effectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8287)
  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            "Unpublished"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_HES2
FEATURES             Location/Qualifiers
     source          1..8287
                     /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(3148..3172)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3225..3266
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3290..3878
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3343..3363)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3761..3772)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3893..4209
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4060..4195
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4257..4304
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4323..4721
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4808..5041
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5113..5234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5150..5169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5204..5223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5274..5409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5429..5448)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5430..5448)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5458..5475)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5458..5474)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5467..5489)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5616..6071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5703..5722)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5913..5934
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6097..6201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6202..6270,6271..7062)
                     /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(6420..6439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7233..7821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7722..7741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7975..7992
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8109..8130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8145..8150,8151..8168,8169..8175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8183..8199
                     /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     8188..8210
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8207..8223
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8207..8223
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8242..8262
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8244..8262
                     /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 ccatggggct gcctcgccgg gcaggggacg
     2641 cggcggagct gcgcaagagc ctgaagccgc tgctggagaa gcgccggcgc gcgcgcatca
     2701 accagagcct gagccagctt aaggggctca tcctgccgct gctgggccgg gagaactcca
     2761 actgctcgaa gctagagaag gcagacgtcc tggaaatgac cgtgcgcttc ctgcaggagc
     2821 tgcctgcgtc ctcatggccc acggcagcgc ccctgccttg cgacagctac cgcgagggct
     2881 acagcgcctg tgtggcgcgc ctggcccgcg tgctgcccgc ctgccgtgtc ctggagcccg
     2941 ccgtgagcgc gcgcctgctg gagcacctgt ggcggagagc ggccagcgcc accctggacg
     3001 gcgggcgcgc tggggattcc agtggcccgt ctgcccccgc cccagcgccc gcgtctgccc
     3061 cagagcccgc atccgctccg gtgccctcgc cgccctcgcc tccctgcggc cctggcctct
     3121 ggcggccgtg gtagaagggc gaattcgacc cagctttctt gtacaaagtg gttgatatcc
     3181 agcacagtgg cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta
     3241 accctctcct cggtctcgat tctacgcgta ccggttagta atgatcgaca atcaacctct
     3301 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3361 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3421 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3481 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3541 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3601 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3661 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3721 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3781 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3841 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggcgatggta ccggtgtgga
     3901 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3961 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4081 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4141 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4201 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     4261 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     4321 ccatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     4381 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     4441 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg
     4501 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     4561 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc
     4621 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     4681 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agcacaattc gagctcggta
     4741 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     4801 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     4861 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4921 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4981 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5041 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     5101 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5161 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5221 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     5281 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5341 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5401 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     5461 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     5521 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5581 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     5641 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5701 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5761 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5821 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5881 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5941 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6001 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6061 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6121 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6181 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6241 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6301 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6361 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6421 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6481 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6541 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6601 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6661 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6721 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6781 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6841 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6901 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6961 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7021 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7081 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7141 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7201 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7261 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7321 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7381 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7441 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7501 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7561 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7621 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7681 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7741 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7801 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7861 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7921 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7981 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8041 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8101 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8161 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8221 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
     8281 caagctt
//
