LOCUS       sequence-471863-        5491 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express NanoLuc(R)-hCHMP4B 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 5491)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5491)
  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: NanoLuc-hCHMP4B Vector
FEATURES             Location/Qualifiers
     source          1..5491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(994..1506)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(1545..1563)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1546..1563)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1683..1986
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2141..2161
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2194..2241
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2273..3067
                     /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(2327..2346)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2937..2956
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3225..3813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3305..3324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3939..3987
                     /note="synthetic polyadenylation signal"
     CDS             complement(4011..5048)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4024..4032
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(5079..5436)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5093..5228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5245..5436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5393..5413
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 catggatcca gcccagttct ccaattcctt catgtcgtcg tcctcctctt ctttcttctt
      361 ggcgggtttt gatggtaggg ctatagaggg aacatttggt agagggactg tttcgggtcc
      421 actgatttcc agcaaattct tgtctagttc ctcctgttct agttcttcta attccgccat
      481 gagctcatcc tcgtcaaact cttctccaaa ccctacaggt ttcgaaattg ctgttgaaat
      541 ctcctctgca agttcttgct ggtcagcaat gtcctgcatt aactcatcaa ctttatcgat
      601 gtccatgttg tcatgggccg ccttcatggc cttggcggca tagcccatgt tcttgagcac
      661 ctcggtgttg gtgttggcat tctccagggc ctcccgctgg aactcgatgg ttgataatgt
      721 gccgtcgatc tgcgccagct gcttctcata cctcttctta cgcttcagtg cctggagggc
      781 cgcgcgcttg tttttggtgc cgtgcttctt ggcggccgtc agctcctgct cgattttctt
      841 ctccaggaac tcctgtttct tgcttaacat ctcttccgtg tcccgcagcc gctggatggc
      901 ctcctggggg gtcgggccgc ccttgccggc cttaccccct ccagccccga acagcttccc
      961 gaacaccgac atggcgatcg cgccgctcga gcccgccaga atgcgttcgc acagccgcca
     1021 gccggtcact ccgttgatgg ttactcggaa cagcagggag ccgtcggggt tgatcaggcg
     1081 ctcgtcgata attttgttgc cgttccacag ggtccctgtt acagtgatct ttttgccgtc
     1141 gaacacggcg atgccttcat acggccgtcc gaaatagtcg atcatgttcg gcgtaacccc
     1201 gtcgattacc agtgtgccat agtgcaggat caccttaaag tgatgatcat ccacagggta
     1261 caccacctta aaaatttttt cgatctggcc catttggtcg ccgctcagac cttcatacgg
     1321 gatgatgaca tggatgtcga tcttcagccc attttcaccg ctcaggacaa tcctttggat
     1381 cggagttacg gacaccccga gattctgaaa caaactggac acacctccct gttcaaggac
     1441 ttggtccagg ttgtagccgg ctgtctgtcg ccagtcccca acgaaatctt cgagtgtgaa
     1501 gaccatggtg agcgctagct gcaaaaagaa caagtagctt gtattctata gtgtcaccta
     1561 aatgggcgat ctgacggttc actaaaccag ctctgcttat ataggcctcc ctggcgcgcc
     1621 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     1681 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1741 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     1801 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     1861 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1921 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1981 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     2041 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     2101 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     2161 gtacggtggg aggtcaaatc gaattctaat actgttgaca attaatcatc ggcatagtat
     2221 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga
     2281 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     2341 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2401 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     2461 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     2521 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2581 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2641 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     2701 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     2761 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     2821 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     2881 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     2941 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3001 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3061 cttctgattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     3121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3661 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3781 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     3841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3901 aggatcttca cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag
     3961 atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt ctaaactcga
     4021 cctacctcct tagcgcgcgg tcgtgtactg ggccggcggt tgccgctgtc ggccagcacc
     4081 tcgacgcagc cgtcggtcca tacggctgcg ctccggcgag cgatttgtgt gcgaccgacg
     4141 gtgccggccc cgctgcggac gatggcatcg cagcggcctt gagcccaggc agcatcgtcg
     4201 aagttgccgt ccacgaggct ctggtagagc tgatccaggc cgatgcgcag catgtaggct
     4261 cgcagacgag ggctgccggc cagctcggga tgccggcgct cgaagtagcg agtctgctgc
     4321 tccatgcaag ccagccaggg ccgccagaag aagatgttgg ccacctcgta ctgactgtcc
     4381 ccgaacatag cttcggacca gtcgattacg gcggtgatgc ggccgttgtc ggtcaggacg
     4441 ttgttgctgc cgaagtcggc atggacgagg tggcgcacct cgggacagtc ttcggcccac
     4501 agcatcagtt cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc catcacggtc
     4561 tgccagtggt agacatgggg atcagcaatg gcgcaaatga aatcccgcca ggttgtgtac
     4621 tggccgatgc cttggggccc gaatgggccg aagccgctgg tttgactgag gtcggcggcg
     4681 gcaatagcat ccatagcttc ggcgacaggc tgtaacacag caggcagctc tgtttcgggg
     4741 aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag gctctcgctg
     4801 aactcgccga tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa gtggcggtac
     4861 acgtagcggt ctttgtagaa gccatcagcg cagctattca cccgcagtac atagccgcgt
     4921 ccgccgacat cgaagctgaa ggctcggctc tcttcgccct ccgacaactg catcaggtcg
     4981 ctcacactgt cgaacttctc gatgagaaat ttttcaacgc tggtagcggt gagttcgggc
     5041 ttcttcatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca
     5101 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     5161 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     5221 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     5281 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     5341 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     5401 tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt
     5461 tcagaggtta tttcaggcca tggtgctgcg c
//
