LOCUS       sequence-473379-        5226 bp    DNA     circular SYN 17-JUN-2025
DEFINITION  Express IL-2 in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5226)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5226)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pNL  [NlucP/IL-2/Hygro) Vector
FEATURES             Location/Qualifiers
     source          1..5226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    145..174
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     regulatory      488..497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             494..1006
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             1010..1129
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1181..1302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1192..1211)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1246..1265
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1496..1853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1496..1687
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1519..1539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1704..1839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1766..1785
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1884..2921
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2900..2908
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    2945..2993
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3149..3166)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3320..3908)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3400..3419)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4108..4899,4900..4968))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    5073..5121
                     /note="synthetic polyadenylation signal"
     misc_feature    5135..5226
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5175..5194
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct ttattctttt catctgttta ctcttgctct tgtccaccac
       61 aatatgctat tcacatgttc agtgtagttt tatgacaaag aaaattttct gagttacttt
      121 tgtatcccca cccccttaaa gaaaggagga aaaactgttt catacagaag gcgttaattg
      181 catgaattag agctatcacc taagtgtggg ctaatgtaac aaagagggat ttcacctaca
      241 tccattcagt cagtctttgg gggtttaaag aaattccaaa gagtcatcag aagaggaaaa
      301 atgaaggtaa tgttttttca gacaggtaaa gtctttgaaa atatgtgtaa tatgtaaaac
      361 attttgacac ccccataata tttttccaga attaacagta taaattgcat ctcttgttca
      421 agagttccct atcactctct ttaatcacta ctcacagtaa gcttggcaat ccggtactgt
      481 tggtaaagcc accatggtct tcacactcga agatttcgtt ggggactggc gacagacagc
      541 cggctacaac ctggaccaag tccttgaaca gggaggtgtg tccagtttgt ttcagaatct
      601 cggggtgtcc gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat
      661 cgacatccat gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg gccagatcga
      721 aaaaattttt aaggtggtgt accctgtgga tgatcatcac tttaaggtga tcctgcacta
      781 tggcacactg gtaatcgacg gggttacgcc gaacatgatc gactatttcg gacggccgta
      841 tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgtggaacgg
      901 caacaaaatt atcgacgagc gcctgatcaa ccccgacggc tccctgctgt tccgagtaac
      961 catcaacgga gtgaccggct ggcggctgtg cgaacgcatt ctggcgaatt ctcacggctt
     1021 ccctcccgag gtggaggagc aggccgccgg caccctgccc atgagctgcg cccaggagag
     1081 cggcatggat agacaccctg ctgcttgcgc cagcgccagg atcaacgtct aaggccgcga
     1141 ctctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     1201 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1261 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1321 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1381 tacaaatgtg gtaaaatcga taaggatccg tttgcgtatt gggcgctctt ccgctgatct
     1441 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttagct accttctgag
     1501 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     1561 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     1621 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1681 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     1741 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     1801 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcg
     1861 attcttctga cactagcgcc accatgaaga agcccgaact caccgctacc agcgttgaaa
     1921 aatttctcat cgagaagttc gacagtgtga gcgacctgat gcagttgtcg gagggcgaag
     1981 agagccgagc cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg gtgaatagct
     2041 gcgctgatgg cttctacaaa gaccgctacg tgtaccgcca cttcgccagc gctgcactac
     2101 ccatccccga agtgttggac atcggcgagt tcagcgagag cctgacatac tgcatcagta
     2161 gacgcgccca aggcgttact ctccaagacc tccccgaaac agagctgcct gctgtgttac
     2221 agcctgtcgc cgaagctatg gatgctattg ccgccgccga cctcagtcaa accagcggct
     2281 tcggcccatt cgggccccaa ggcatcggcc agtacacaac ctggcgggat ttcatttgcg
     2341 ccattgctga tccccatgtc taccactggc agaccgtgat ggacgacacc gtgtccgcca
     2401 gcgtagctca agccctggac gaactgatgc tgtgggccga agactgtccc gaggtgcgcc
     2461 acctcgtcca tgccgacttc ggcagcaaca acgtcctgac cgacaacggc cgcatcaccg
     2521 ccgtaatcga ctggtccgaa gctatgttcg gggacagtca gtacgaggtg gccaacatct
     2581 tcttctggcg gccctggctg gcttgcatgg agcagcagac tcgctacttc gagcgccggc
     2641 atcccgagct ggccggcagc cctcgtctgc gagcctacat gctgcgcatc ggcctggatc
     2701 agctctacca gagcctcgtg gacggcaact tcgacgatgc tgcctgggct caaggccgct
     2761 gcgatgccat cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc gctcgccgga
     2821 gcgcagccgt atggaccgac ggctgcgtcg aggtgctggc cgacagcggc aaccgccggc
     2881 ccagtacacg accgcgcgct aaggaggtag gtcgagttta aactctagaa ccggtcatgg
     2941 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgttcgaac
     3001 tagatgctgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     3061 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     3121 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3181 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3241 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3301 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3361 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3421 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3481 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3541 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3601 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3661 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3721 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3781 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3841 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3961 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4021 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagcg
     4081 gccgcaaatg ctaaaccact gcagtggtta ccagtgcttg atcagtgagg caccgatctc
     4141 agcgatctgc ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt agatcactac
     4201 gattcgtgag ggcttaccat caggccccag cgcagcaatg atgccgcgag agccgcgttc
     4261 accggccccc gatttgtcag caatgaacca gccagcaggg agggccgagc gaagaagtgg
     4321 tcctgctact ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag
     4381 aagttcgcca gtgagtagtt tccgaagagt tgtggccatt gctactggca tcgtggtatc
     4441 acgctcgtcg ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac
     4501 atgatcaccc atattatgaa gaaatgcagt cagctcctta gggcctccga tcgttgtcag
     4561 aagtaagttg gccgcggtgt tgtcgctcat ggtaatggca gcactacaca attctcttac
     4621 cgtcatgcca tccgtaagat gcttttccgt gaccggcgag tactcaacca agtcgttttg
     4681 tgagtagtgt atacggcgac caagctgctc ttgcccggcg tctatacggg acaacaccgc
     4741 gccacatagc agtactttga aagtgctcat catcgggaat cgttcttcgg ggcggaaaga
     4801 ctcaaggatc ttgccgctat tgagatccag ttcgatatag cccactcttg cacccagttg
     4861 atcttcagca tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa
     4921 tgccgcaaag aagggaatga gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt
     4981 tcaatattat tgaagcattt atcagggtta ctagtacgtc tctcaaggat aagtaagtaa
     5041 tattaaggta cgggaggtat tggacaggcc gcaataaaat atctttattt tcattacatc
     5101 tgtgtgttgg ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa
     5161 cgaaacaaaa caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat
     5221 ttctct
//
