LOCUS       sequence-472961-        6363 bp    DNA     circular SYN 15-MAY-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 6363)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/IL-2/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6363
                     /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..2143
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2147..2266
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2318..2439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2329..2348)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2383..2402
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2633..2990
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2633..2824
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2656..2676)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2841..2976
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2903..2922
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3021..4058
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4037..4045
                     /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    4082..4130
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4286..4303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4457..5045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4537..4556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5245..6036,6037..6105))
                     /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    6210..6258
                     /note="synthetic polyadenylation signal"
     misc_feature    6272..6363
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6312..6331
                     /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 accatggaag atgccaaaaa cattaagaag ggcccagcgc cattctaccc
      541 actcgaagac gggaccgccg gcgagcagct gcacaaagcc atgaagcgct acgccctggt
      601 gcccggcacc atcgccttta ccgacgcaca tatcgaggtg gacattacct acgccgagta
      661 cttcgagatg agcgttcggc tggcagaagc tatgaagcgc tatgggctga atacaaacca
      721 tcggatcgtg gtgtgcagcg agaatagctt gcagttcttc atgcccgtgt tgggtgccct
      781 gttcatcggt gtggctgtgg ccccagctaa cgacatctac aacgagcgcg agctgctgaa
      841 cagcatgggc atcagccagc ccaccgtcgt attcgtgagc aagaaagggc tgcaaaagat
      901 cctcaacgtg caaaagaagc taccgatcat acaaaagatc atcatcatgg atagcaagac
      961 cgactaccag ggcttccaaa gcatgtacac cttcgtgact tcccatttgc cacccggctt
     1021 caacgagtac gacttcgtgc ccgagagctt cgaccgggac aaaaccatcg ccctgatcat
     1081 gaacagtagt ggcagtaccg gattgcccaa gggcgtagcc ctaccgcacc gcaccgcttg
     1141 tgtccgattc agtcatgccc gcgaccccat cttcggcaac cagatcatcc ccgacaccgc
     1201 tatcctcagc gtggtgccat ttcaccacgg cttcggcatg ttcaccacgc tgggctactt
     1261 gatctgcggc tttcgggtcg tgctcatgta ccgcttcgag gaggagctat tcttgcgcag
     1321 cttgcaagac tataagattc aatctgccct gctggtgccc acactattta gcttcttcgc
     1381 taagagcact ctcatcgaca agtacgacct aagcaacttg cacgagatcg ccagcggcgg
     1441 ggcgccgctc agcaaggagg taggtgaggc cgtggccaaa cgcttccacc taccaggcat
     1501 ccgccagggc tacggcctga cagaaacaac cagcgccatt ctgatcaccc ccgaagggga
     1561 cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc gaggctaagg tggtggactt
     1621 ggacaccggt aagacactgg gtgtgaacca gcgcggcgag ctgtgcgtcc gtggccccat
     1681 gatcatgagc ggctacgtta acaaccccga ggctacaaac gctctcatcg acaaggacgg
     1741 ctggctgcac agcggcgaca tcgcctactg ggacgaggac gagcacttct tcatcgtgga
     1801 ccggctgaag agcctgatca aatacaaggg ctaccaggta gccccagccg aactggagag
     1861 catcctgctg caacacccca acatcttcga cgccggggtc gccggcctgc ccgacgacga
     1921 tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca tgaccgagaa
     1981 ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc aagaagctgc gcggtggtgt
     2041 tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag ttggacgccc gcaagatccg
     2101 cgagattctc attaaggcca agaagggcgg caagatcgcc gtgaattctc acggcttccc
     2161 tcccgaggtg gaggagcagg ccgccggcac cctgcccatg agctgcgccc aggagagcgg
     2221 catggataga caccctgctg cttgcgccag cgccaggatc aacgtctaag gccgcgactc
     2281 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     2341 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2401 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2461 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     2521 aaatgtggta aaatcgataa ggatccgttt gcgtattggg cgctcttccg ctgatctgcg
     2581 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttagctacc ttctgaggcg
     2641 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2701 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2761 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2821 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2881 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2941 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcgatt
     3001 cttctgacac tagcgccacc atgaagaagc ccgaactcac cgctaccagc gttgaaaaat
     3061 ttctcatcga gaagttcgac agtgtgagcg acctgatgca gttgtcggag ggcgaagaga
     3121 gccgagcctt cagcttcgat gtcggcggac gcggctatgt actgcgggtg aatagctgcg
     3181 ctgatggctt ctacaaagac cgctacgtgt accgccactt cgccagcgct gcactaccca
     3241 tccccgaagt gttggacatc ggcgagttca gcgagagcct gacatactgc atcagtagac
     3301 gcgcccaagg cgttactctc caagacctcc ccgaaacaga gctgcctgct gtgttacagc
     3361 ctgtcgccga agctatggat gctattgccg ccgccgacct cagtcaaacc agcggcttcg
     3421 gcccattcgg gccccaaggc atcggccagt acacaacctg gcgggatttc atttgcgcca
     3481 ttgctgatcc ccatgtctac cactggcaga ccgtgatgga cgacaccgtg tccgccagcg
     3541 tagctcaagc cctggacgaa ctgatgctgt gggccgaaga ctgtcccgag gtgcgccacc
     3601 tcgtccatgc cgacttcggc agcaacaacg tcctgaccga caacggccgc atcaccgccg
     3661 taatcgactg gtccgaagct atgttcgggg acagtcagta cgaggtggcc aacatcttct
     3721 tctggcggcc ctggctggct tgcatggagc agcagactcg ctacttcgag cgccggcatc
     3781 ccgagctggc cggcagccct cgtctgcgag cctacatgct gcgcatcggc ctggatcagc
     3841 tctaccagag cctcgtggac ggcaacttcg acgatgctgc ctgggctcaa ggccgctgcg
     3901 atgccatcgt ccgcagcggg gccggcaccg tcggtcgcac acaaatcgct cgccggagcg
     3961 cagccgtatg gaccgacggc tgcgtcgagg tgctggccga cagcggcaac cgccggccca
     4021 gtacacgacc gcgcgctaag gaggtaggtc gagtttaaac tctagaaccg gtcatggccg
     4081 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg ttcgaactag
     4141 atgctgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     4201 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     4261 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4321 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4381 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4441 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4501 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4561 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4621 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4681 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4741 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4801 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4861 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4921 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4981 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5041 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5101 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5161 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc
     5221 gcaaatgcta aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc
     5281 gatctgccta tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat
     5341 tcgtgagggc ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc
     5401 ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc
     5461 tgctactttg tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag
     5521 ttcgccagtg agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg
     5581 ctcgtcgttc ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg
     5641 atcacccata ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag
     5701 taagttggcc gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt
     5761 catgccatcc gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga
     5821 gtagtgtata cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc
     5881 acatagcagt actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc
     5941 aaggatcttg ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc
     6001 ttcagcatct tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc
     6061 cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca
     6121 atattattga agcatttatc agggttacta gtacgtctct caaggataag taagtaatat
     6181 taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt
     6241 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
     6301 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
     6361 tct
//
