LOCUS       sequence-479804-        5123 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-TCEB2 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 5123)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5123)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-TCEB2 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5123
                     /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(2066..2084)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2121..2242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2132..2151)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2186..2205
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2487..2844
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2487..2678
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2510..2530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2695..2830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2859..2906
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2938..3732
                     /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(2992..3011)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3602..3621
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3796..3844
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4083..4671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4163..4182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4787..5070
                     /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 ggacgtgttc ctcatgatcc
     1621 ggcgccacaa gaccaccatc ttcacggacg ccaaggagtc cagcacggtg ttcgaactga
     1681 agcgcatcgt cgagggcatc ctcaagcggc ctcctgacga gcagcggctg tacaaggatg
     1741 accaactctt ggatgatggc aagacactgg gcgagtgtgg cttcaccagt caaacagcac
     1801 ggccacaggc cccagccaca gtggggctgg ccttccgggc agatgacacc tttgaggccc
     1861 tgtgcatcga gccgttttcc agcccgccag agctgcccga tgtgatgaag ccccaggact
     1921 cgggaagcag tgccaatgaa caagccgtgc aggtttaaac gaattcgggc tcggtacccg
     1981 gggatcctct agagtcgacc tgcaggcatg caagctgatc cggctgctaa caaagcccga
     2041 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcg
     2101 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2161 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2221 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2281 gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     2341 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     2401 gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct
     2461 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     2521 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2581 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2641 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2761 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2821 tttggaggcc taggcttttg caaaaagctt aattaactgt tgacaattaa tcatcggcat
     2881 agtatatcgg catagtataa tacgacaagg tgaggaacta aacccaggag gcagatcatg
     2941 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3001 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3061 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3121 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3181 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3241 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3301 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3361 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3421 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3481 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3541 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3601 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3661 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3721 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3781 catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     3841 tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc
     3901 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacccggta
     3961 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4021 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4081 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4141 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4201 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4261 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4321 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4381 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4441 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4501 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4561 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4621 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct
     4681 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4741 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg aaatacagga
     4801 acgcacgctg gatggccctt cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg
     4861 gattaagtgg gggtaatgtg gcctgtaccc tctggttgca taggtattca tacggttaaa
     4921 atttatcagg cgcgattgcg gcagtttttc gggtggtttg ttgccatttt tacctgtctg
     4981 ctgccgtgat cgcgctgaac gcgttttagc ggtgcgtaca attaagggat tatggtaaat
     5041 ccacttactg tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg
     5101 tccggcgtag aggatcgaga tct
//
