LOCUS       sequence-476608-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express AKT3-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 6044)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6044)
  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            "Unpublished"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: AKT3-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2523..3035
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3123..3141)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3178..3299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3189..3208)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3243..3262
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3544..3901
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3544..3735
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3567..3587)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3752..3887
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3916..3963
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3995..4789
                     /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(4049..4068)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4659..4678
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4853..4901
                     /note="synthetic polyadenylation signal"
     rep_origin      5058..5603
                     /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     5504..5523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5720..5737
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatgagc gatgttacca
     1081 ttgtgaaaga aggttgggtt cagaagaggg gagaatatat aaaaaactgg aggccaagat
     1141 acttcctttt gaagacagat ggctcattca taggatataa agagaaacct caagatgtgg
     1201 atttacctta tcccctcaac aacttttcag tggcaaaatg ccagttaatg aaaacagaac
     1261 gaccaaagcc aaacacattt ataatcagat gtctccagtg gactactgtt atagagagaa
     1321 catttcatgt agatactcca gaggaaaggg aagaatggac agaagctatc caggctgtag
     1381 cagacagact gcagaggcaa gaagaggaga gaatgaattg tagtccaact tcacaaattg
     1441 ataatatagg agaggaagag atggatgcct ctacaaccca tcataaaaga aagacaatga
     1501 atgattttga ctatttgaaa ctactaggta aaggcacttt tgggaaagtt attttggttc
     1561 gagagaaggc aagtggaaaa tactatgcta tgaagattct gaagaaagaa gtcattattg
     1621 caaaggatga agtggcacac actctaactg aaagcagagt attaaagaac actagacatc
     1681 cctttttaac atccttgaaa tattccttcc agacaaaaga ccgtttgtgt tttgtgatgg
     1741 aatatgttaa tgggggcgag ctgtttttcc atttgtcgag agagcgggtg ttctctgagg
     1801 accgcacacg tttctatggt gcagaaattg tctctgcctt ggactatcta cattccggaa
     1861 agattgtgta ccgtgatctc aagttggaga atctaatgct ggacaaagat ggccacataa
     1921 aaattacaga ttttggactt tgcaaagaag ggatcacaga tgcagccacc atgaagacat
     1981 tctgtggcac tccagaatat ctggcaccag aggtgttaga agataatgac tatggccgag
     2041 cagtagactg gtggggccta ggggttgtca tgtatgaaat gatgtgtggg aggttacctt
     2101 tctacaacca ggaccatgag aaactttttg aattaatatt aatggaagac attaaatttc
     2161 ctcgaacact ctcttcagat gcaaaatcat tgctttcagg gctcttgata aaggatccaa
     2221 ataaacgcct tggtggagga ccagatgatg caaaagaaat tatgagacac agtttcttct
     2281 ctggagtaaa ctggcaagat gtatatgata aaaagcttgt acctcctttt aaacctcaag
     2341 taacatctga gacagatact agatattttg atgaagaatt tacagctcag actattacaa
     2401 taacaccacc tgaaaaatat gatgaggatg gtatggactg catggacaat gagaggcggc
     2461 cgcatttccc tcaattttcc tactctgcaa gtggacgaga agtttctctc ggctcgagcg
     2521 gcgtcttcac actcgaagat ttcgttgggg actggcgaca gacagccggc tacaacctgg
     2581 accaagtcct tgaacaggga ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa
     2641 ctccgatcca aaggattgtc ctgagcggtg aaaatgggct gaagatcgac atccatgtca
     2701 tcatcccgta tgaaggtctg agcggcgacc aaatgggcca gatcgaaaaa atttttaagg
     2761 tggtgtaccc tgtggatgat catcacttta aggtgatcct gcactatggc acactggtaa
     2821 tcgacggggt tacgccgaac atgatcgact atttcggacg gccgtatgaa ggcatcgccg
     2881 tgttcgacgg caaaaagatc actgtaacag ggaccctgtg gaacggcaac aaaattatcg
     2941 acgagcgcct gatcaacccc gacggctccc tgctgttccg agtaaccatc aacggagtga
     3001 ccggctggcg gctgtgcgaa cgcattctgg cgtaaggccg cgactctaga gtcgacctgc
     3061 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     3121 caccgctgag caataactag cataacccct tggggcggcc gcttcgagca gacatgataa
     3181 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3241 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3301 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     3361 taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt
     3421 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3481 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3541 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3721 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3841 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3901 aaagcttaat taactgttga caattaatca tcggcatagt atatcggcat agtataatac
     3961 gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc
     4021 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4081 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4141 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4201 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4261 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4321 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4381 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4441 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4501 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4561 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4621 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4681 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4741 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4801 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat
     4861 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat
     4921 ccgctcatgc gatgataagc tgtcaaacat gagaattaca acttatatcg tatggggctg
     4981 acttcaggtg ctacatttga agagataaat tgcactgaaa tctagaaata ttttatctga
     5041 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
     5101 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
     5161 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
     5221 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
     5281 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5341 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
     5401 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
     5461 accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac gcctggtatc
     5521 tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg tgatgcttgt
     5581 caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt ccctgttaag
     5641 tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg
     5701 cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata tcctgtatca
     5761 catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag cacttcactg
     5821 acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc tgatgtccgg
     5881 cggtgctttt gccgttacgc accaccccgt catgagatta cgcgttttag cggtgcgtac
     5941 aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg agataaaccg
     6001 cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
