LOCUS       sequence-470865-        6236 bp    DNA     circular SYN 01-MAY-2025
DEFINITION  Express hIL8 response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6236)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6236)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL4 [luc2P/hIL8/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      361..370
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             367..2016
                     /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             2020..2139
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2191..2312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2202..2221)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2256..2275
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2506..2863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2506..2697
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2529..2549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2714..2849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2776..2795
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2894..3931
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3910..3918
                     /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    3955..4003
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4159..4176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4330..4918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4410..4429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5118..5909,5910..5978))
                     /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    6083..6131
                     /note="synthetic polyadenylation signal"
     misc_feature    6145..6236
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6185..6204
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct ctagaaataa aaaagcatac aattgataat tcaccaaatt
       61 gtggagcttc agtattttaa atgtatatta aaattaaatt attttaaaga tcaaagaaaa
      121 ctttcgtcat actccgtatt tgataaggaa caaataggaa gtgtgatgac tcaggtttgc
      181 cctgagggga tgggccatca gttgcaaatc gtggaatttc ctctgacata atgaaaagat
      241 gagggtgcat aagttctcta gtagggtgat gatataaaaa gccaccggag cactccataa
      301 ggcacaaact ttcagagaca gcagagcaca caagcttggc aatccggtac tgttggtaaa
      361 gccaccatgg aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa
      421 gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc
      481 accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag
      541 atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc
      601 gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc
      661 ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg
      721 ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac
      781 gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac
      841 cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag
      901 tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt
      961 agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga
     1021 ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc
     1081 agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc
     1141 ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa
     1201 gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc
     1261 actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg
     1321 ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag
     1381 ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag
     1441 cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc
     1501 ggtaagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg
     1561 agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg
     1621 cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg
     1681 aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg
     1741 ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc
     1801 gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc
     1861 gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc
     1921 gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt
     1981 ctcattaagg ccaagaaggg cggcaagatc gccgtgaatt ctcacggctt ccctcccgag
     2041 gtggaggagc aggccgccgg caccctgccc atgagctgcg cccaggagag cggcatggat
     2101 agacaccctg ctgcttgcgc cagcgccagg atcaacgtct aaggccgcga ctctagagtc
     2161 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2221 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2281 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2341 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2401 gtaaaatcga taaggatccg tttgcgtatt gggcgctctt ccgctgatct gcgcagcacc
     2461 atggcctgaa ataacctctg aaagaggaac ttggttagct accttctgag gcggaaagaa
     2521 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2581 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcg attcttctga
     2881 cactagcgcc accatgaaga agcccgaact caccgctacc agcgttgaaa aatttctcat
     2941 cgagaagttc gacagtgtga gcgacctgat gcagttgtcg gagggcgaag agagccgagc
     3001 cttcagcttc gatgtcggcg gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg
     3061 cttctacaaa gaccgctacg tgtaccgcca cttcgccagc gctgcactac ccatccccga
     3121 agtgttggac atcggcgagt tcagcgagag cctgacatac tgcatcagta gacgcgccca
     3181 aggcgttact ctccaagacc tccccgaaac agagctgcct gctgtgttac agcctgtcgc
     3241 cgaagctatg gatgctattg ccgccgccga cctcagtcaa accagcggct tcggcccatt
     3301 cgggccccaa ggcatcggcc agtacacaac ctggcgggat ttcatttgcg ccattgctga
     3361 tccccatgtc taccactggc agaccgtgat ggacgacacc gtgtccgcca gcgtagctca
     3421 agccctggac gaactgatgc tgtgggccga agactgtccc gaggtgcgcc acctcgtcca
     3481 tgccgacttc ggcagcaaca acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga
     3541 ctggtccgaa gctatgttcg gggacagtca gtacgaggtg gccaacatct tcttctggcg
     3601 gccctggctg gcttgcatgg agcagcagac tcgctacttc gagcgccggc atcccgagct
     3661 ggccggcagc cctcgtctgc gagcctacat gctgcgcatc ggcctggatc agctctacca
     3721 gagcctcgtg gacggcaact tcgacgatgc tgcctgggct caaggccgct gcgatgccat
     3781 cgtccgcagc ggggccggca ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt
     3841 atggaccgac ggctgcgtcg aggtgctggc cgacagcggc aaccgccggc ccagtacacg
     3901 accgcgcgct aaggaggtag gtcgagttta aactctagaa ccggtcatgg ccgcaataaa
     3961 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgttcgaac tagatgctgt
     4021 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     4081 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     4141 gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4741 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagcg gccgcaaatg
     5101 ctaaaccact gcagtggtta ccagtgcttg atcagtgagg caccgatctc agcgatctgc
     5161 ctatttcgtt cgtccatagt ggcctgactc cccgtcgtgt agatcactac gattcgtgag
     5221 ggcttaccat caggccccag cgcagcaatg atgccgcgag agccgcgttc accggccccc
     5281 gatttgtcag caatgaacca gccagcaggg agggccgagc gaagaagtgg tcctgctact
     5341 ttgtccgcct ccatccagtc tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca
     5401 gtgagtagtt tccgaagagt tgtggccatt gctactggca tcgtggtatc acgctcgtcg
     5461 ttcggtatgg cttcgttcaa ctctggttcc cagcggtcaa gccgggtcac atgatcaccc
     5521 atattatgaa gaaatgcagt cagctcctta gggcctccga tcgttgtcag aagtaagttg
     5581 gccgcggtgt tgtcgctcat ggtaatggca gcactacaca attctcttac cgtcatgcca
     5641 tccgtaagat gcttttccgt gaccggcgag tactcaacca agtcgttttg tgagtagtgt
     5701 atacggcgac caagctgctc ttgcccggcg tctatacggg acaacaccgc gccacatagc
     5761 agtactttga aagtgctcat catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc
     5821 ttgccgctat tgagatccag ttcgatatag cccactcttg cacccagttg atcttcagca
     5881 tcttttactt tcaccagcgt ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag
     5941 aagggaatga gtgcgacacg aaaatgttgg atgctcatac tcgtcctttt tcaatattat
     6001 tgaagcattt atcagggtta ctagtacgtc tctcaaggat aagtaagtaa tattaaggta
     6061 cgggaggtat tggacaggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     6121 ttttttgtgt gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa
     6181 caaactagca aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctct
//
