LOCUS       sequence-450400-        8893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription factor EZH2 with a specific barcode assigned..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8893)
  TITLE     The Immune Top TF library is a collection of Immune-related human TF
            ORFs in LV vectors. For each TF, an 8 nt seq is assigned, allowing 
            simple identification during single-cell RNA-seq.
REFERENCE   2  (bases 1 to 8893)
  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"
COMMENT     Sequence Label: SFFV-EZH2-Brd 228
FEATURES             Location/Qualifiers
     source          1..8893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(93..112)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(94..112)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(122..139)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(122..138)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(131..153)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      280..735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(367..386)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     577..598
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        761..865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(866..934,935..1726)
                     /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(1084..1103)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1897..2485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2386..2405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2639..2656
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2773..2794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2809..2814,2815..2832,2833..2839)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2847..2863
                     /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     2852..2874
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2871..2887
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2871..2887
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2906..2926
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2908..2926
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2954..3180
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3181..3361
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3408..3533
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4026..4259
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4444..4488
                     /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    4755..4872
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4906..5312
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    7843..8246
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             8365..8598
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8670..8791
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8707..8726)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8761..8780
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      join(8831..8893,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
       61 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
      121 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
      181 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
      241 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
      301 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      361 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      421 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      481 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      541 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      601 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      661 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      721 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
      781 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      841 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      901 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      961 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1021 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1081 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1141 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1201 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1261 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1321 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1381 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1441 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1501 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1561 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1621 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1681 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1741 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1801 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1861 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1921 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1981 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2041 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2101 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2161 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2221 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2281 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2341 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2401 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2461 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2521 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2581 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2641 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2701 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     2761 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     2821 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2881 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     2941 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
     3001 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     3061 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
     3121 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
     3181 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     3241 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     3301 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     3361 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     3421 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     3481 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     3541 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     3601 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     3661 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     3721 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     3781 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     3841 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     3901 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     3961 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     4021 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     4081 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     4141 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     4201 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     4261 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     4321 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     4381 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     4441 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     4501 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     4561 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     4621 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     4681 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     4741 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     4801 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     4861 attcaaaatt ttatcgatca cgagactagc ctcgagctgg ctgcagtaac gccattttgc
     4921 aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc gggtacatga
     4981 aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc cccggcccgg
     5041 ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga acagatggtc
     5101 cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt ccaggctccc
     5161 ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct gcttctcgct
     5221 tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc ctcactcggc
     5281 gcgccagtcc tccgacagac tgagtcggcc ggtcgaatca agcttatcga taccgtcgac
     5341 ggatccgcta gcccgcggtt cgaaatgggc cagactggga agaaatctga gaagggacca
     5401 gtttgttggc ggaagcgtgt aaaatcagag tacatgcgac tgagacagct caagaggttc
     5461 agacgagctg atgaagtaaa gagtatgttt agttccaatc gtcagaaaat tttggaaaga
     5521 acggaaatct taaaccaaga atggaaacag cgaaggatac agcctgtgca catcctgact
     5581 tctgtgagct cattgcgcgg gactagggag tgttcggtga ccagtgactt ggattttcca
     5641 acacaagtca tcccattaaa gactctgaat gcagttgctt cagtacccat aatgtattct
     5701 tggtctcccc tacagcagaa ttttatggtg gaagatgaaa ctgttttaca taacattcct
     5761 tatatgggag atgaagtttt agatcaggat ggtactttca ttgaagaact aataaaaaat
     5821 tatgatggga aagtacacgg ggatagagaa tgtgggttta taaatgatga aatttttgtg
     5881 gagttggtga atgcccttgg tcaatataat gatgatgacg atgatgatga tggagacgat
     5941 cctgaagaaa gagaagaaaa gcagaaagat ctggaggatc accgagatga taaagaaagc
     6001 cgcccacctc ggaaatttcc ttctgataaa atttttgaag ccatttcctc aatgtttcca
     6061 gataagggca cagcagaaga actaaaggaa aaatataaag aactcaccga acagcagctc
     6121 ccaggcgcac ttcctcctga atgtaccccc aacatagatg gaccaaatgc taaatctgtt
     6181 cagagagagc aaagcttaca ctcctttcat acgcttttct gtaggcgatg ttttaaatat
     6241 gactgcttcc tacatcgtaa gtgcaattat tcttttcatg caacacccaa cacttataag
     6301 cggaagaaca cagaaacagc tctagacaac aaaccttgtg gaccacagtg ttaccagcat
     6361 ttggagggag caaaggagtt tgctgctgct ctcaccgctg agcggataaa gaccccacca
     6421 aaacgtccag gaggccgcag aagaggacgg cttcccaata acagtagcag gcccagcacc
     6481 cccaccatta atgtgctgga atcaaaggat acagacagtg atagggaagc agggactgaa
     6541 acggggggag agaacaatga taaagaagaa gaagagaaga aagatgaaac ttcgagctcc
     6601 tctgaagcaa attctcggtg tcaaacacca ataaagatga agccaaatat tgaacctcct
     6661 gagaatgtgg agtggagtgg tgctgaagcc tcaatgttta gagtcctcat tggcacttac
     6721 tatgacaatt tctgtgccat tgctaggtta attgggacca aaacatgtag acaggtgtat
     6781 gagtttagag tcaaagaatc tagcatcata gctccagctc ccgctgagga tgtggatact
     6841 cctccaagga aaaagaagag gaaacaccgg ttgtgggctg cacactgcag aaagatacag
     6901 ctgaaaaagg acggctcctc taaccatgtt tacaactatc aaccctgtga tcatccacgg
     6961 cagccttgtg acagttcgtg cccttgtgtg atagcacaaa atttttgtga aaagttttgt
     7021 caatgtagtt cagagtgtca aaaccgcttt ccgggatgcc gctgcaaagc acagtgcaac
     7081 accaagcagt gcccgtgcta cctggctgtc cgagagtgtg accctgacct ctgtcttact
     7141 tgtggagccg ctgaccattg ggacagtaaa aatgtgtcct gcaagaactg cagtattcag
     7201 cggggctcca aaaagcatct attgctggca ccatctgacg tggcaggctg ggggattttt
     7261 atcaaagatc ctgtgcagaa aaatgaattc atctcagaat actgtggaga gattatttct
     7321 caagatgaag ctgacagaag agggaaagtg tatgataaat acatgtgcag ctttctgttc
     7381 aacttgaaca atgattttgt ggtggatgca acccgcaagg gtaacaaaat tcgttttgca
     7441 aatcattcgg taaatccaaa ctgctatgca aaagttatga tggttaacgg tgatcacagg
     7501 ataggtattt ttgccaagag agccatccag actggcgaag agctgttttt tgattacaga
     7561 tacagccagg ctgatgccct gaagtatgtc ggcatcgaaa gagaaatgga aatcccttga
     7621 ccggtcggat cccccgggct gcaggaattc gagcatctta ccgccattta ttcccatatt
     7681 tgttctgttt ttcttgattt gggtatacat ttaaatgtta ataaaacaaa atggtggggc
     7741 aatcatttac atttttaggg atatgtaatt actagttcag gtgtattgcc acaagacaaa
     7801 catgttaaga aactttcccg ttatttacgc tctgttcctg ttaatcaacc tctggattac
     7861 aaaatttgtg aaagattgac tgatattctt aactatgttg ctccttttac gctgtgtgga
     7921 tatgctgctt taatgcctct gtatcatgct attgcttccc gtacggcttt cgttttctcc
     7981 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtccgtcaa
     8041 cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca ctggctgggg cattgccacc
     8101 acctgtcaac tcctttctgg gactttcgct ttccccctcc cgatcgccac ggcagaactc
     8161 atcgccgcct gccttgcccg ctgctggaca ggggctaggt tgctgggcac tgataattcc
     8221 gtggtgttgt cggggaagct gacgtccttt gcaattcgat atcaagctgt accttctaga
     8281 tgtacactaa tccaagcgct tctagaccaa tgacttacaa ggcagctgta gatcttagcc
     8341 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     8401 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8461 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8521 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8581 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     8641 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     8701 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     8761 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     8821 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     8881 actaattttt ttt
//
