LOCUS       sequence-450421-        7179 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Transcription factor NFE4 with a specific barcode assigned..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7179)
  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 7179)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SFFV-NFE4-Brd 247
FEATURES             Location/Qualifiers
     source          1..7179
                     /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..736
                     /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        762..866
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(867..935,936..1727)
                     /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(1085..1104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1898..2486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2387..2406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2640..2657
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2774..2795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2810..2815,2816..2833,2834..2840)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2848..2864
                     /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     2853..2875
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2872..2888
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2872..2888
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2907..2927
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2909..2927
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2955..3181
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3182..3362
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3409..3534
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4027..4260
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4445..4489
                     /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    4756..4873
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4907..5313
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    6129..6532
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             6651..6884
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6956..7077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6993..7012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7047..7066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      join(7117..7179,1..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 aaaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      721 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      781 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      841 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      901 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      961 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1021 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1081 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1141 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1201 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1261 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1321 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1381 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1441 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1501 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1561 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1621 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1681 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1741 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1801 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1861 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1921 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1981 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2041 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2101 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2161 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2221 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2281 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2341 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2401 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2461 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2521 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2581 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2641 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     2701 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     2761 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     2821 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     2881 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     2941 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     3001 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     3061 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     3121 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     3181 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     3241 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     3301 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     3361 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     3421 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     3481 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     3541 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     3601 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     3661 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     3721 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     3781 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     3841 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     3901 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     3961 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     4021 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     4081 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     4141 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     4201 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     4261 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     4321 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     4381 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     4441 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     4501 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     4561 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     4621 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     4681 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
     4741 acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     4801 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     4861 aattcaaaat tttatcgatc acgagactag cctcgagctg gctgcagtaa cgccattttg
     4921 caaggcatgg aaaaatacca aaccaagaat agagaagttc agatcaaggg cgggtacatg
     4981 aaaatagcta acgttgggcc aaacaggata tctgcggtga gcagtttcgg ccccggcccg
     5041 gggccaagaa cagatggtca ccgcagtttc ggccccggcc cgaggccaag aacagatggt
     5101 ccccagatat ggcccaaccc tcagcagttt cttaagaccc atcagatgtt tccaggctcc
     5161 cccaaggacc tgaaatgacc ctgcgcctta tttgaattaa ccaatcagcc tgcttctcgc
     5221 ttctgttcgc gcgcttctgc ttcccgagct ctataaaaga gctcacaacc cctcactcgg
     5281 cgcgccagtc ctccgacaga ctgagtcggc cggtcgaatc aagcttatcg ataccgtcga
     5341 cggatccgct agcccgcggt tcgaactgcc tcgtgttgtc tgttggcaca ctctcaagag
     5401 tttgaacgga tacaagaatc tttcatctgg tgccgaaacc cgggaggggc tccggtcttc
     5461 gtcccccgtg gacctacccc tccgccccag aaagcaggcc acagcagccg gacaaaggaa
     5521 gctcctcagc ctccagttgc ttctctgtgc atgcacatca gtcactgatc tcacctactg
     5581 gggccctgca ggccatgggg ccacagctcc acacagaagc ctcctagcaa tccacctcca
     5641 cctggtgcct gcttcaagtg cggcaatgaa ggccactggc ccacacaatg cccaaaccca
     5701 ggtaaaccca cgaggccatg ccccctctgc ggaggacccc actggaagtt ggactgtgag
     5761 cggcccctgc aaggaccacc cccatccctt cctgagccaa tcaaaccctc ctactcggat
     5821 ctcgtcagcc ttgccgctga agactgatag tgccttggaa cagacacccc agcaactacc
     5881 atcgcttcat ctgagccaag ggtagaccgg tcggatcccc cgggctgcag gaattcgagc
     5941 atcttaccgc catttattcc catatttgtt ctgtttttct tgatttgggt atacatttaa
     6001 atgttaataa aacaaaatgg tggggcaatc atttacattt ttagggatat gtaattacta
     6061 gttcaggtgt attgccacaa gacaaacatg ttaagaaact ttcccgttat ttacgctctg
     6121 ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag attgactgat attcttaact
     6181 atgttgctcc ttttacgctg tgtggatatg ctgctttaat gcctctgtat catgctattg
     6241 cttcccgtac ggctttcgtt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     6301 aggagttgtg gcccgttgtc cgtcaacgtg gcgtggtgtg ctctgtgttt gctgacgcaa
     6361 cccccactgg ctggggcatt gccaccacct gtcaactcct ttctgggact ttcgctttcc
     6421 ccctcccgat cgccacggca gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6481 ctaggttgct gggcactgat aattccgtgg tgttgtcggg gaagctgacg tcctttgcaa
     6541 ttcgatatca agctgtacct tctagatgta cagcgtactt agcgcttcta gaccaatgac
     6601 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     6661 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     6721 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6781 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6841 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     6901 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6961 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     7021 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     7081 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     7141 tccgcccatt ctccgcccca tggctgacta atttttttt
//
