LOCUS       sequence-450424-        7528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transcription factor LEF1 with a specific barcode assigned..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7528)
  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 7528)
  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-LEF1-Brd 196
FEATURES             Location/Qualifiers
     source          1..7528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /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    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1960..2366
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      2395..2404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3532..3935
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             4054..4287
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4359..4480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4396..4415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4450..4469
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4520..4655
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4675..4694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4676..4694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4704..4721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4704..4720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4713..4735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4862..5317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4949..4968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5159..5180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5343..5447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5448..5516,5517..6308)
                     /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(5666..5685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6479..7067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6968..6987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7221..7238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7355..7376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7391..7396,7397..7414,7415..7421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7429..7445
                     /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     7434..7456
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7453..7469
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7453..7469
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7488..7508
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7490..7508
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttatcg atcacgagac tagcctcgag ctggctgcag taacgccatt ttgcaaggca
     1981 tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag
     2041 ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa
     2101 gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga
     2161 tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg
     2221 acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt
     2281 cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact cggcgcgcca
     2341 gtcctccgac agactgagtc ggccggtcga atcaagctta tcgataccgt cgacgccacc
     2401 atggccgaag tggctagaca ggcccagaca agccaagagc cttaccacga caaggccaga
     2461 gagcaccctg acgacggcaa acatcctgat ggcggcctgt acaacaaggg ccctagctac
     2521 agcagctact ccggctatat catgatgccc aacatgaaca acgaccccta catgagcaac
     2581 ggcagcctgt ctcctcctat tcctcggacc agcaacaagg tgccagtggt gcagccttct
     2641 cacgccgtgc atcctctgac acccctgatc acctacagcg acgagcactt cagccctggc
     2701 tctcaccctt ctcacatccc cagcgacgtg aacagcaagc agggcatgag cagacaccct
     2761 ccagctccag acatccccac attctaccca ctgtctccag gcggcgtggg ccagattaca
     2821 cctccactcg gatggttcag ccaccacatg atccctggac cacctggacc tcacaccacc
     2881 ggcattcctc atcctgccat cgtgacccct caagtgaagc aagagcaccc acacaccgac
     2941 agcgacctga tgcacgtgaa gcctcagcac gagcagcgga aagagcaaga gcctaagcgg
     3001 ccccacatca agaagcccct gaacgccttc atgctgtaca tgaaagaaat gcgggccaac
     3061 gtggtggccg agtgtaccct gaaagagagc gccgccatta accagatcct gggcagaaga
     3121 tggcacgccc tgagcagaga agaacaggcc aagtactacg agctggcccg gaaagaacgg
     3181 cagctgcaca tgcaactgta ccccggctgg agcgccagag acaactacgg caagaagaag
     3241 aagcgcaagc gcgagaagct gcaagagtct gcctctggca caggccctag aatgacagcc
     3301 gcctacatct gataaggatc ccccgggctg caggaattcg agcatcttac cgccatttat
     3361 tcccatattt gttctgtttt tcttgatttg ggtatacatt taaatgttaa taaaacaaaa
     3421 tggtggggca atcatttaca tttttaggga tatgtaatta ctagttcagg tgtattgcca
     3481 caagacaaac atgttaagaa actttcccgt tatttacgct ctgttcctgt taatcaacct
     3541 ctggattaca aaatttgtga aagattgact gatattctta actatgttgc tccttttacg
     3601 ctgtgtggat atgctgcttt aatgcctctg tatcatgcta ttgcttcccg tacggctttc
     3661 gttttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3721 gtccgtcaac gtggcgtggt gtgctctgtg tttgctgacg caacccccac tggctggggc
     3781 attgccacca cctgtcaact cctttctggg actttcgctt tccccctccc gatcgccacg
     3841 gcagaactca tcgccgcctg ccttgcccgc tgctggacag gggctaggtt gctgggcact
     3901 gataattccg tggtgttgtc ggggaagctg acgtcctttg caattcgata tcaagctgta
     3961 ccttctagat gtacagatta ttaagcgctt ctagaccaat gacttacaag gcagctgtag
     4021 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     4081 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     4141 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     4201 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     4261 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     4321 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     4381 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4441 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     4501 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4561 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4621 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     4681 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5281 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     5341 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5401 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5461 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5521 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5581 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5641 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5701 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5761 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5821 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5881 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5941 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6001 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6061 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6121 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6181 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6241 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6301 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6361 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6421 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7081 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7141 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7201 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     7261 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     7321 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     7381 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     7441 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     7501 actaaaggga acaaaagctg gagctgca
//
