LOCUS       sequence-472459-        7055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PI4KB-NanoLuc(R) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7055)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7055)
  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            "Unpublished"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: PI4KB-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1095..1114)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1096..1114)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1156..1175)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          1158..1290
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(1426..1629)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1455..1475)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1630..2009
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(2454..2471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2588..3133)
                     /direction=LEFT
                     /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     complement(2668..2687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3290..3338
                     /note="synthetic polyadenylation signal"
     CDS             complement(3402..4196)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3513..3532)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4123..4142
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4228..4275)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4290..4647)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4304..4439
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4456..4647
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4604..4624
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4892..5013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4929..4948)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4983..5002
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5050..5068
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5156..5668)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
ORIGIN
        1 tggttcagca gagacagctc ggagatcagc cgctgtgtct tctgctcctt tgtgggcagg
       61 gtggccaggc gcttgccgat tgccatcagg gacttgataa actccctctc tggtgccagt
      121 ctcactgggg agctgaagga attgtcgatg ctctcggtgc tgctgctcag ctcctcatcc
      181 tcattctcca ccttagggtt gctggctgtc ctcttcaggt tagaggacag agagatggag
      241 gcggtggcgt cgctcttaga cctctggtgt gttctcttgg atgggctcag tccggtatca
      301 ggtgctggag acagggatgg cagctccctc ttcctgtgtg caggcttcag ctcgtcgctc
      361 aggatcagct ttctcagctt tgtgccccgg ctgtggcgct gggtagagat gtgcatatcg
      421 ctagaatagg cgcccagcag cagggcacac tgcagagaaa aattgatgga ctgcctgcat
      481 ctgtgcacga tgtaaggctt gatggcgtcg cccacatcct cgtccatgtg gatatacatg
      541 ttcagcagct ggggcaggta gaaatccacg tcctcattcc gaaagcagaa caggcggttg
      601 ccgatgtagg cctgcacgcc aggctccttg gaattataca ggtagctgat ggccatagag
      661 atgtcaaaca gcttgctctc gaacagccgc agcagccagg actgcttggc gctattgttc
      721 tggcgccttc tccgggcgcc ctttgctgtt ccggatgcca ctgctgctcc catctcatcc
      781 tcctcctccc tgatctgggc gggtggatcg tccagacatc tgatctcgct atccactccg
      841 tcgccgttca ccagctccag tggggttccc ctgctgctca cggccactcc tccgtgcagc
      901 agcttcacct tctccagcac ctcctggcag gccttctggg ccacctcagg gtcgatcacg
      961 ctcagctctc ccactccctc tgtgatcacg ctcagcagag agccgccatt gttgccaggg
     1021 gggccagatg ttggctcgga ggtaggcttc agtggggcag gctccaccac ggtatcgccc
     1081 atggcgatcg ctagccctat agtgagtcgt attaatactc tagccttaag agctgtaatt
     1141 gaactgggag tggacacctg tggagagaaa ggcaaagtgg atgtcagtaa gaccaatagg
     1201 tgcctatcag aaacgcaaga gtcttctctg tctcgacaag cccagtttct attggtctcc
     1261 ttaaacctgt cttgtaacct tgatacttac ctgcccagtg cctcacgacc aacttctgca
     1321 gcttaagttc gagactgttg tgtcagaagc actgactgcg ttagcaattt aactgtgata
     1381 aactaccgca ataaagcttc tagtgatctg acggttcact aaaccagctc tgcttatata
     1441 gacctcccac cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt tattacgaca
     1501 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     1561 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattggtgt actgccaaaa
     1621 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     1681 ccgtaaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     1741 agggggcgga cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     1801 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     1861 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     1921 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     1981 ctattaataa ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat
     2041 ataaatgtac atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat
     2101 ttatgctata taaccaatga ataatatggc taatggccaa tattgaagat ctcgatcctc
     2161 tacgccggac gcatcgtggc cggagtactg cggtttatct cgatggctac gagggcagac
     2221 agtaagtgga tttaccataa tcccttaatt gtacgcaccg ctaaaacgcg taatctcatg
     2281 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
     2341 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
     2401 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
     2461 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
     2521 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
     2581 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
     2641 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
     2701 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     2761 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     2821 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     2881 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     2941 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     3001 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     3061 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     3121 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     3181 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     3241 atgtttgaca gcttatcatc gcatgagcgg atccttatcg ctatcgattc acacaaaaaa
     3301 ccaacacaca gatgtaatga aaataaagat attttattgc ggccatcgtg atggcaggtt
     3361 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     3421 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     3481 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     3541 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     3601 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     3661 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     3721 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     3781 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     3841 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     3901 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     3961 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     4021 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     4081 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     4141 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgatc
     4201 tgcctcctgg gtttagttcc tcaccttgtc gtattatact atgccgatat actatgccga
     4261 tgattaattg tcaacagtta attaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     4321 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     4381 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     4441 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     4501 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     4561 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     4621 ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct ttcagaggtt
     4681 atttcaggcc atggtgctgc gcagatccgc gtatgcggtg tgaaataccg cacagatgcg
     4741 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgattt
     4801 taccacattt gtagaggttt tacttgctta aaaaaacctc ccacatctcc ccctgaacct
     4861 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4921 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4981 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc ggccgcccca
     5041 aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct
     5101 ttgttagcag ccggatcagc ttgcatgcct gcaggtcgac tctagagtcg cggccttacg
     5161 ccagaatgcg ttcgcacagc cgccagccgg tcactccgtt gatggttact cggaacagca
     5221 gggagccgtc ggggttgatc aggcgctcgt cgataatttt gttgccgttc cacagggtcc
     5281 ctgttacagt gatctttttg ccgtcgaaca cggcgatgcc ttcatacggc cgtccgaaat
     5341 agtcgatcat gttcggcgta accccgtcga ttaccagtgt gccatagtgc aggatcacct
     5401 taaagtgatg atcatccaca gggtacacca ccttaaaaat tttttcgatc tggcccattt
     5461 ggtcgccgct cagaccttca tacgggatga tgacatggat gtcgatcttc agcccatttt
     5521 caccgctcag gacaatcctt tggatcggag ttacggacac cccgagattc tgaaacaaac
     5581 tggacacacc tccctgttca aggacttggt ccaggttgta gccggctgtc tgtcgccagt
     5641 ccccaacgaa atcttcgagt gtgaagacgc cgctcgagcc gagagaaacc atgatgccat
     5701 tggtcagata ctgaaagccg tcgtacagct ttgtggtgat gctccgcata gagccatcca
     5761 ccatctgctc caccagcagc tgcagctgct cctctgtcat ggacatgtga aaccgctcct
     5821 tcaggttgcg gatggtgcta gagccgtgga aacagggcag ctggctgccc tgctgcatga
     5881 tctccacgat ctgcaccacc ttgtccatgt gcttccttgc tgcgatcagt ccctgcagca
     5941 tcagcatctt gtagtagttg aacatatcgc cgtccaggcc gcccatcaca tccacgaact
     6001 ctgtggtcag cttaaaggcg ctggtctcga agcccaggtt gcgtggggag ctagacagga
     6061 taaagccgaa gtcgatgtgg atgatgtggc cctcggcatc cagcaggata ttgccgttgt
     6121 gcctgtcctt cacctgcagc agatagcaca ccaggcagta gccggcgcag ctctgcacaa
     6181 aatttctctg ggcagacagg aaggcctctg tggtatagga gccgtgctcc tgcaggaagt
     6241 agtccagcag ggacagctgg ctctgcttct tcacctggtg gatggacacg gcgttcacca
     6301 cgggctcgat catgccagaa tcggcggaga tgaccaggat cttataaggc ttgatccaca
     6361 ggggcacccg ctcctgctcc cagatgctct gcagctgctt cagcacctga aatgccagca
     6421 gctcctgcct cagatcgtcg ccacacttca cgatcacaga cagcagcctc cagtttggca
     6481 ggtggccgta aggggagccc tctctgatcc ggcgcacctt ctcctgccat ggctccttca
     6541 gggccacggc agatggatcc tctgggtccc tcttgaaggc tgtaggggtg tgggccagct
     6601 gctcggacag ccttctccgg atgtcgcctg ctgcgataaa cacgggctcc ttgctctcct
     6661 gagatgtgat ggaatccacg ctgaactgag agatattgtc gcagctgttg gtgtgcacct
     6721 ctggcagctc cacctgcagc tcgccgatat cgtccacaga ccaggcctca tcgtcattat
     6781 cgtagttggg cactgtggag aagctgccgg ctctctgctc gtgggtgatg ccacactctg
     6841 gcagattctc cacggaccgg gtgctcctga ttctgttctc agggatgcgg gcgggcacgg
     6901 atgtggtgtc aaaattctcg cactccagca cctccacata gatcaggtaa ggggccttat
     6961 ccttagagtt cagcaccacg gcctgtgtgt ggggcaccct caccacgtgg tggtcgaatc
     7021 ctgcggttgg cagccacact cttgctggca gcttg
//
