LOCUS       sequence-473195-        8519 bp    DNA     circular SYN 20-MAY-2025
DEFINITION  Transcription factor NR4A3 with a specific barcode assigned..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8519)
  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 8519)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SFFV-NR4A3-Brd 28
FEATURES             Location/Qualifiers
     source          1..8519
                     /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"
     CDS             5647..5676
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5650..5679
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5653..5682
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5656..5685
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5659..5688
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    7469..7872
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             7991..8224
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8296..8417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8333..8352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8387..8406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      join(8457..8519,1..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 cgaaatgcca tgtgttcaag cacagtactc cccctcacca
     5401 cccggcagct cctacgcagc acaaacctat tccagtgagt atacaacaga aattatgaat
     5461 ccggattata caaaacttac gatggattta gggtccacag aaataactgc gacggctacg
     5521 acttcattgc catctattag cacatttgtc gaagggtatt catccaatta tgagttaaaa
     5581 ccctcttgtg tctatcagat gcaacgacca ctgattaagg ttgaagaagg ccgcgcacct
     5641 tcctatcacc atcaccatca tcatcatcat catcatcatc atcaccatca acaacaacac
     5701 caacaaccca gcattccccc tgcttcatct cccgaagatg aagttcttcc ttctacatct
     5761 atgtatttta aacaatctcc gcctagcaca ccaactacac ccgcgtttcc acctcaagca
     5821 ggcgcactct gggatgaagc cttgccatca gcccctgggt gtattgcccc aggacctctt
     5881 ttggatccac ccatgaaagc agtaccaact gtcgctggtg cccgttttcc acttttccat
     5941 tttaaaccat caccccctca cccgccagca ccttctcctg ctggcggaca tcatctggga
     6001 tatgatccca ccgcggcagc tgcactgagt ttacctcttg gggcagcggc agctgcagga
     6061 tctcaagcag ctgctctgga atctcatccc tatgggctcc cattagcaaa acgtgccgca
     6121 cctcttgcat ttcccccgct gggtcttacc cctagtccca cggcaagttc acttcttggg
     6181 gaatctccat ctctccccag tcccccatct cgttcaagta gttcagggga agggacatgc
     6241 gctgtttgtg gagataatgc tgcgtgtcaa cattatggtg ttcggacttg tgaaggatgt
     6301 aaagggttct ttaaacgaac ggttcaaaag aacgccaagt acgtgtgtct cgctaacaag
     6361 aattgtcctg ttgataaacg ccgcagaaat cggtgccaat attgcagatt ccaaaagtgc
     6421 ctgtctgtcg ggatggtcaa agaggtggtg cggaccgact ccctcaaggg acggcgcggg
     6481 cgactcccct ctaaacccaa atctcccttg cagcaagagc catcccaacc aagcccgcca
     6541 agtcccccta tttgtatgat gaacgcactg gtgagagccc ttaccgattc cactccacgg
     6601 gacttggact actctcggta ttgccctacc gatcaagcgg cagccggaac tgacgcagaa
     6661 cacgtccagc agttttataa tcttttgacc gcgagtatcg acgttagccg ctcatgggct
     6721 gagaaaatcc ccgggttcac cgaccttcca aaagaggacc aaaccttgct gatagagagc
     6781 gctttcctgg aattattcgt tctgcggctg tctattagat ccaatacagc cgaggacaaa
     6841 ttcgttttct gtaacgggtt ggtactccac cgcttgcaat gtttgagggg cttcggagaa
     6901 tggctggata gcataaagga tttcagtctc aacctgcaaa gtctcaatct ggacattcag
     6961 gcactcgctt gtttgagcgc gctctctatg attaccgagc gccacggact gaaggagccc
     7021 aaacgggtag aggaattatg taataaaata acgtcatccc tcaaggatca tcaatctaaa
     7081 gggcaagcgc tcgaacctac agaaagcaaa gtgctcggag ctttagtcga gttgcgcaaa
     7141 atttgtacac tcggtcttca acggatcttt tatctcaaac tcgaggatct tgtctccccg
     7201 ccatctatta tcgataaact gtttctggat acacttccct tttagaccgg tcggatcccc
     7261 cgggctgcag gaattcgagc atcttaccgc catttattcc catatttgtt ctgtttttct
     7321 tgatttgggt atacatttaa atgttaataa aacaaaatgg tggggcaatc atttacattt
     7381 ttagggatat gtaattacta gttcaggtgt attgccacaa gacaaacatg ttaagaaact
     7441 ttcccgttat ttacgctctg ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag
     7501 attgactgat attcttaact atgttgctcc ttttacgctg tgtggatatg ctgctttaat
     7561 gcctctgtat catgctattg cttcccgtac ggctttcgtt ttctcctcct tgtataaatc
     7621 ctggttgctg tctctttatg aggagttgtg gcccgttgtc cgtcaacgtg gcgtggtgtg
     7681 ctctgtgttt gctgacgcaa cccccactgg ctggggcatt gccaccacct gtcaactcct
     7741 ttctgggact ttcgctttcc ccctcccgat cgccacggca gaactcatcg ccgcctgcct
     7801 tgcccgctgc tggacagggg ctaggttgct gggcactgat aattccgtgg tgttgtcggg
     7861 gaagctgacg tcctttgcaa ttcgatatca agctgtacct tctagatgta cacaaaaact
     7921 agcgcttcta gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     7981 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     8041 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     8101 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     8161 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     8221 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     8281 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     8341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     8401 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     8461 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttt
//
