LOCUS       sequence-471881-        6491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-ACT1 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 6491)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6491)
  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            "Unpublished"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: NanoLuc-ACT1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6491
                     /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             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3434..3452)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3489..3610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3500..3519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3554..3573
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3855..4212
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3855..4046
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3878..3898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4063..4198
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4227..4274
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4306..5100
                     /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(4360..4379)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4970..4989
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5164..5212
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5451..6039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5531..5550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6155..6438
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat gcctcctcag cttcaagaaa
     1621 ctagaatgaa ccgaagcatt cctgtggagg ttgatgaatc agaaccatac ccaagtcagt
     1681 tgctgaaacc aatcccagaa tattccccgg aagaggaatc agaaccacct gctccaaata
     1741 taaggaacat ggcacccaac agcttgtctg cacccacaat gcttcacaat tcctccggag
     1801 acttttctca agctcactca accctgaaac ttgcaaatca ccagcggcct gtatcccggc
     1861 aggtcacctg cctgcgcact caagttctgg aggacagtga agacagtttc tgcaggagac
     1921 acccaggcct gggcaaagct ttcccttctg ggtgctctgc agtcagcgag cctgcgtctg
     1981 agtctgtggt tggagccctc cctgcagagc atcagttttc atttatggaa aaacgtaatc
     2041 aatggctggt atctcagctt tcagcggctt ctcctgacac tggccatgac tcagacaaat
     2101 cagaccaaag tttacctaat gcctcagcag actccttggg cggtagccag gagatggtgc
     2161 aacggcccca gcctcacagg aaccgagcag gcctggatct gccaaccata gacacgggat
     2221 atgattccca gccccaggat gtcctgggca tcaggcagct ggaaaggccc ctgcccctca
     2281 cctccgtgtg ttacccccag gacctcccca gacctctcag gtccagggag ttccctcagt
     2341 ttgaacctca gaggtatcca gcatgtgcac agatgctgcc tcccaatctt tccccacatg
     2401 ctccatggaa ctatcattac cattgtcctg gaagtcccga tcaccaggtg ccatatggcc
     2461 atgactaccc tcgagcagcc taccagcaag tgatccagcc ggctctgcct gggcagcccc
     2521 tgcctggagc cagtgtgaga ggcctgcacc ctgtgcagaa ggttatcctg aattatccca
     2581 gcccctggga ccaagaagag aggcccgcac agagagactg ctcctttccg gggcttccaa
     2641 ggcaccagga ccagccacat caccagccac ctaatagagc tggtgctcct ggggagtcct
     2701 tggagtgccc tgcagagctg agaccacagg ttccccagcc tccgtcccca gctgctgtgc
     2761 ctagaccccc tagcaaccct ccagccagag gaactctaaa aacaagcaat ttgccagaag
     2821 aattgcggaa agtctttatc acttattcga tggacacagc tatggaggtg gtgaaattcg
     2881 tgaacttttt gttggtaaat ggcttccaaa ctgcaattga catatttgag gatagaatcc
     2941 gaggcattga tatcattaaa tggatggagc gctaccttag ggataagacc gtgatgataa
     3001 tcgtagcaat cagccccaaa tacaaacagg acgtggaagg cgctgagtcg cagctggacg
     3061 aggatgagca tggcttacat actaagtaca ttcatcgaat gatgcagatt gagttcataa
     3121 aacaaggaag catgaatttc agattcatcc ctgtgctctt cccaaatgct aagaaggagc
     3181 atgtgcccac ctggcttcag aacactcatg tctacagctg gcccaagaat aaaaaaaaca
     3241 tcctgctgcg gctgctgaga gaggaagagt atgtggctcc tccacggggg cctctgccca
     3301 cccttcaggt ggttcccttg gtttaaacga attcgggctc ggtacccggg gatcctctag
     3361 agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag
     3421 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc
     3481 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3541 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3601 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3661 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa
     3721 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3781 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     3841 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4021 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4081 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4141 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4201 ggcttttgca aaaagcttaa ttaactgttg acaattaatc atcggcatag tatatcggca
     4261 tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat
     4321 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4381 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4441 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4501 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4561 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4621 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4681 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4741 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4801 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4861 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4921 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4981 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5041 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5101 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     5161 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     5221 gcgataagga tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     5281 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acccggtatc agctcactca
     5341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5881 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6001 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     6061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6121 tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac gcacgctgga
     6181 tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga ttaagtgggg
     6241 gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat ttatcaggcg
     6301 cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct gccgtgatcg
     6361 cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc
     6421 tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag
     6481 gatcgagatc t
//
