LOCUS       sequence-471880-        6465 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express ACT1-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 6465)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6465)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: ACT1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6465
                     /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             2808..3320
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3408..3426)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3463..3584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3474..3493)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3528..3547
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3829..4186
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3829..4020
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3852..3872)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4037..4172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4201..4248
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4280..5074
                     /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(4334..4353)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4944..4963
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5138..5186
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5425..6013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5505..5524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6129..6412
                     /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 ggctagcgat cgccatgcct cctcagcttc
     1081 aagaaactag aatgaaccga agcattcctg tggaggttga tgaatcagaa ccatacccaa
     1141 gtcagttgct gaaaccaatc ccagaatatt ccccggaaga ggaatcagaa ccacctgctc
     1201 caaatataag gaacatggca cccaacagct tgtctgcacc cacaatgctt cacaattcct
     1261 ccggagactt ttctcaagct cactcaaccc tgaaacttgc aaatcaccag cggcctgtat
     1321 cccggcaggt cacctgcctg cgcactcaag ttctggagga cagtgaagac agtttctgca
     1381 ggagacaccc aggcctgggc aaagctttcc cttctgggtg ctctgcagtc agcgagcctg
     1441 cgtctgagtc tgtggttgga gccctccctg cagagcatca gttttcattt atggaaaaac
     1501 gtaatcaatg gctggtatct cagctttcag cggcttctcc tgacactggc catgactcag
     1561 acaaatcaga ccaaagttta cctaatgcct cagcagactc cttgggcggt agccaggaga
     1621 tggtgcaacg gccccagcct cacaggaacc gagcaggcct ggatctgcca accatagaca
     1681 cgggatatga ttcccagccc caggatgtcc tgggcatcag gcagctggaa aggcccctgc
     1741 ccctcacctc cgtgtgttac ccccaggacc tccccagacc tctcaggtcc agggagttcc
     1801 ctcagtttga acctcagagg tatccagcat gtgcacagat gctgcctccc aatctttccc
     1861 cacatgctcc atggaactat cattaccatt gtcctggaag tcccgatcac caggtgccat
     1921 atggccatga ctaccctcga gcagcctacc agcaagtgat ccagccggct ctgcctgggc
     1981 agcccctgcc tggagccagt gtgagaggcc tgcaccctgt gcagaaggtt atcctgaatt
     2041 atcccagccc ctgggaccaa gaagagaggc ccgcacagag agactgctcc tttccggggc
     2101 ttccaaggca ccaggaccag ccacatcacc agccacctaa tagagctggt gctcctgggg
     2161 agtccttgga gtgccctgca gagctgagac cacaggttcc ccagcctccg tccccagctg
     2221 ctgtgcctag accccctagc aaccctccag ccagaggaac tctaaaaaca agcaatttgc
     2281 cagaagaatt gcggaaagtc tttatcactt attcgatgga cacagctatg gaggtggtga
     2341 aattcgtgaa ctttttgttg gtaaatggct tccaaactgc aattgacata tttgaggata
     2401 gaatccgagg cattgatatc attaaatgga tggagcgcta ccttagggat aagaccgtga
     2461 tgataatcgt agcaatcagc cccaaataca aacaggacgt ggaaggcgct gagtcgcagc
     2521 tggacgagga tgagcatggc ttacatacta agtacattca tcgaatgatg cagattgagt
     2581 tcataaaaca aggaagcatg aatttcagat tcatccctgt gctcttccca aatgctaaga
     2641 aggagcatgt gcccacctgg cttcagaaca ctcatgtcta cagctggccc aagaataaaa
     2701 aaaacatcct gctgcggctg ctgagagagg aagagtatgt ggctcctcca cgggggcctc
     2761 tgcccaccct tcaggtggtt cccttggttt ctctcggctc gagcggcgtc ttcacactcg
     2821 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     2881 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     2941 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     3001 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     3061 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     3121 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     3181 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     3241 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     3301 gcgaacgcat tctggcgtaa ggccgcgact ctagagtcga cctgcaggca tgcaagctga
     3361 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     3421 actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt
     3481 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3541 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3601 ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc
     3661 tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     3721 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     3781 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     3841 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3961 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4021 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4081 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4141 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact
     4201 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     4261 taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4321 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4381 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4441 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4501 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4561 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4621 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4681 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4741 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4801 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4861 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4921 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4981 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5041 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5101 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     5161 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc
     5221 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     5281 cggactggct ttctacccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5341 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5401 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5461 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5521 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5581 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5641 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5701 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5761 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5821 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     5881 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5941 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6001 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6061 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6121 tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc
     6181 atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg
     6241 cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt
     6301 tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta
     6361 caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc
     6421 gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatct
//
