LOCUS       sequence-470564-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HiBiT-IRAK4 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-IRAK4 Vector
FEATURES             Location/Qualifiers
     source          1..5751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..520
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     440..458
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     primer_bind     525..544
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        525..543
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        549..567
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     549..566
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2147..2165)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2202..2323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2213..2232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2267..2286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2568..2925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2568..2759
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2591..2611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2776..2911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2950..2959
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2956..3354
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3379..3427
                     /note="synthetic polyadenylation signal"
     promoter        3583..3687
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3688..3756,3757..4548)
                     /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(3906..3925)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4765..5310
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5211..5230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5427..5444
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agatctccac ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac
       61 gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat
      121 tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg
      181 cgacgcttcc tcgtccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga
      241 cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag
      301 caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg
      361 ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg
      421 ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag
      481 tcggctccct cgttgaccga atcaccgacc tctctcccca agcttaatac gactcactat
      541 agggtacgat ttaggtgaca ctatagaata agctagcgct caccatggtg agcggctggc
      601 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgaaca
      661 aacccataac accatcaaca tatgtgcgct gcctcaatgt tggactaatt aggaagctgt
      721 cagattttat tgatcctcaa gaaggatgga agaagttagc tgtagctatt aaaaaaccat
      781 ctggtgatga tagatacaat cagtttcaca taaggagatt tgaagcatta cttcaaactg
      841 gaaaaagtcc cacttctgaa ttactgtttg actggggcac cacaaattgc acagttggtg
      901 atcttgtgga tcttttgatc caaaatgaat tttttgctcc tgcaagtctt ttgctcccag
      961 atgctgttcc caaaactgct aatacactac cttctaaaga agctataaca gttcagcaaa
     1021 aacagatgcc tttctgtgac aaagacagga cattgatgac acctgtgcag aatcttgaac
     1081 aaagctatat gccacctgac tcctcaagtc cagaaaataa aagtttagaa gttagtgata
     1141 cacgttttca cagtttttca ttttatgaat tgaagaatgt cacaaataac tttgatgaac
     1201 gacccatttc tgttggtggt aataaaatgg gagagggagg atttggagtt gtatataaag
     1261 gctacgtaaa taacacaact gtggcagtga agaagcttgc agcaatggtt gacattacta
     1321 ctgaagaact gaaacagcag tttgatcaag aaataaaagt aatggcaaag tgtcaacatg
     1381 aaaacttagt agaactactt ggtttctcaa gtgatggaga tgacctctgc ttagtatatg
     1441 tttacatgcc taatggttca ttgctagaca gactctcttg cttggatggt actccaccac
     1501 tttcttggca catgagatgc aagattgctc agggtgcagc taatggcatc aattttctac
     1561 atgaaaatca tcatattcat agagatatta aaagtgcaaa tatcttactg gatgaagctt
     1621 ttactgctaa aatatctgac tttggccttg cacgggcttc tgagaagttt gcccagacag
     1681 tcatgactag cagaattgtg ggaacaacag cttatatggc accagaagct ttgcgtggag
     1741 aaataacacc caaatctgat atttacagct ttggtgtggt tttactagaa ataataactg
     1801 gacttccagc tgtggatgaa caccgtgaac ctcagttatt gctagatatt aaagaagaaa
     1861 ttgaagatga agaaaagaca attgaagatt atattgataa aaagatgaat gatgctgatt
     1921 ccacttcagt tgaagctatg tactctgttg ctagtcaatg tctgcatgaa aagaaaaata
     1981 agagaccaga cattaagaag gttcaacagc tgctgcaaga gatgacagct tctgtttaaa
     2041 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     2101 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2161 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2221 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2281 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2341 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     2401 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     2461 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     2521 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     2581 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2641 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2701 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2761 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2821 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2881 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cgattcttct
     2941 gacactagcg ccaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga
     3001 gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct
     3061 ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct
     3121 tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt
     3181 atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag
     3241 gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg
     3301 acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaatgatct
     3361 agaaccggtc atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     3421 ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata
     3481 gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgta
     3541 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3601 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3661 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3721 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3781 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3841 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cactttcaaa
     3901 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3961 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4021 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4081 gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc ttttttgcac
     4141 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4201 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4261 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4321 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4381 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4441 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4501 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt cgaaatgacc
     4561 gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4621 aatcagggga taacgcagga aagaacatgt caaacatgag aattacaact tatatcgtat
     4681 ggggctgact tcaggtgcta catttgaaga gataaattgc actgaaatct agaaatattt
     4741 tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa tctcttgctc
     4801 tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag ctaccaactc
     4861 tttgaaccga ggtaactggc ttggaggagc gcagtcacca aaacttgtcc tttcagttta
     4921 gccttaaccg gcgcatgact tcaagactaa ctcctctaaa tcaattacca gtggctgctg
     4981 ccagtggtgc ttttgcatgt ctttccgggt tggactcaag acgatagtta ccggataagg
     5041 cgcagcggtc ggactgaacg gggggttcgt gcatacagtc cagcttggag cgaactgcct
     5101 acccggaact gagtgtcagg cgtggaatga gacaaacgcg gccataacag cggaatgaca
     5161 ccggtaaacc gaaaggcagg aacaggagag cgcacgaggg agccgccagg gggaaacgcc
     5221 tggtatcttt atagtcctgt cgggtttcgc caccactgat ttgagcgtca gatttcgtga
     5281 tgcttgtcag gggggcggag cctatggaaa aacggctttg ccgcggccct ctcacttccc
     5341 tgttaagtat cttcctggca tcttccagga aatctccgcc ccgttcgtaa gccatttccg
     5401 ctcgccgcag tcgaacgacc gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc
     5461 tgtatcacat attctgctga cgcaccggtg cagccttttt tctcctgcca catgaagcac
     5521 ttcactgaca ccctcatcag tgccaacata gtaagccagt atacactccg ctagcgctga
     5581 tgtccggcgg tgcttttgcc gttacgcacc accccgtcat gagattacgc gttttagcgg
     5641 tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga
     5701 taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
