LOCUS       sequence-481951-        5904 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express NanoLuc-TCEB1 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 5904)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5904)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-TCEB1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1557..2351
                     /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(1611..1630)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2221..2240
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2375..2423
                     /note="synthetic polyadenylation signal"
     rep_origin      2549..3137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3038..3057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3309..3968)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3883..3902
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4000..4047)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(4080..4100)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4255..4558
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4678..4696
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4678..4695
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4735..5247
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    5807..5888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5844..5863)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg caggcagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa agccgaattc tgcagatatc
     1201 catcacactg tcgagcatgc atctagtcca gtgtggtgga attcggcttc gtggggacgc
     1261 cgtacaggga cgtgcacctc tcccgctgca ccgcctccag cgtcgccgcc ggctcgaagg
     1321 acggggccgg gatgacgatg caggcggcgt gggaggtggc gcccaagttg cccatgacca
     1381 tgccgaagca gtggtagaag ggcaccggca gacacacccg gtcctgctcc gtgtagccga
     1441 ccgtgcggcc cacccagtag ccgttgttga ggatgttgtg gtgggagagc gtggcgccct
     1501 tggggaagcc ggtgtgccgg aggtgtactg gatgttgacc gggaatcgat gccaccatga
     1561 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     1621 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     1681 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     1741 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     1801 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     1861 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     1921 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     1981 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2041 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     2101 aggatctcgt cgtgacccac ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2161 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2221 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2281 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2341 agttcttctg aactctagcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     2401 tctgtgtgtt ggttttttgt gtgacgattc cggactataa aaggatctag gtgaagatcc
     2461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2521 accccgtaga aaagatcaaa ggatcttctt gaaatccttt ttttctgcgc gtaatctgct
     2581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     2701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3241 ccgcctttga gtgagctgat accggttggg cgtcgcttgg tcggtcattt cgaaccccag
     3301 agtcccgctt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     3361 ctgccgacat ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc
     3421 accttgtcgc cttgcgtata atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc
     3481 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     3541 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     3601 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     3661 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     3721 accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca
     3781 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     3841 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     3901 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     3961 ttctccatga tctgcctcct gggtttagtt cctcaccttg tcgtattata ctatgccgat
     4021 atactatgcc gatgattaat tgtcaacagt attagaattc gatttgacct cccaccgtac
     4081 acgcctaccg cccatttgcg tcaacggggc ggggttatta cgacattttg gaaagtcccg
     4141 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     4201 ccgtgagtca aaccgctatc cacgcccatt ggtgtactgc caaaaccgca tcaccatggt
     4261 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccgta aggtcatgta
     4321 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcggacttgg
     4381 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     4441 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     4501 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     4561 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt
     4621 ggcgcgccag ggaggcctat ataagcagag ctggtttagt gaaccgtcag atcgcccatt
     4681 taggtgacac tatagaatac aagctacttg ttctttttgc agctagcgct caccatggtc
     4741 ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa
     4801 gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg
     4861 atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc
     4921 ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg
     4981 taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac
     5041 ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc
     5101 gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag
     5161 cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc
     5221 tggcggctgt gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat ggatggagag
     5281 gagaaaacct atggtggctg tgaaggacct gatgccatgt atgtcaaatt gatatcatct
     5341 gatggccatg aatttattgt aaaaagagaa catgcattaa catcaggcac gataaaagcc
     5401 atgttgagtg gcccaggtca gtttgctgag aacgaaacca atgaggtcaa ttttagagag
     5461 ataccttcac atgtgctatc gaaagtatgc atgtatttta cgtacaaggt tcgctacact
     5521 aacagctcca ccgagattcc tgaattccca attgcacctg aaattgcact ggaactgctg
     5581 atggctgcga acttcttaga ttgtgtttaa acgaattcgg gctcggtacc cggggatcct
     5641 ctagaaacac ctaggctctc gtacgaagtc agctgataac gtcgacgtat tgcggccgcg
     5701 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     5761 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     5821 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5881 ttcactgccg acgtcacgcg tctt
//
