LOCUS       sequence-482027-        6674 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express NanoLuc-Syk 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 6674)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6674)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-Syk Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(169..372)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(198..218)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        373..752
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    943..1226
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1342..1930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1831..1850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2169..2217
                     /note="synthetic polyadenylation signal"
     CDS             complement(2281..3075)
                     /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(2392..2411)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3002..3021
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3107..3154)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3169..3526)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3183..3318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3335..3526
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3483..3503
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    3771..3892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3808..3827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3862..3881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3929..3947
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5987..6499)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(6512..6531)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6513..6531)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6573..6592)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          join(6575..6674,1..33)
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
ORIGIN
        1 tccttaaacc tgtcttgtaa ccttgatact tacctgccca gtgcctcacg accaacttct
       61 gcagcttaag ttcgagactg ttgtgtcaga agcactgact gcgttagcaa tttaactgtg
      121 ataaactacc gcaataaagc ttctagtgat ctgacggttc actaaaccag ctctgcttat
      181 atagacctcc caccgtacac gcctaccgcc catttgcgtc aacggggcgg ggttattacg
      241 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
      301 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattgg tgtactgcca
      361 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
      421 gtcccgtaag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
      481 aatagggggc ggacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
      541 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
      601 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
      661 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
      721 ttactattaa taactagtca ataatcaatg ccaacatggc ggtcatattg gacatgagcc
      781 aatataaatg tacatattat gatatagata caacgtatgc aatggccaat agccaatatt
      841 gatttatgct atataaccaa tgaataatat ggctaatggc caatattgaa gatctcgatc
      901 ctctacgccg gacgcatcgt ggccggagta ctgcggttta tctcgatggc tacgagggca
      961 gacagtaagt ggatttacca taatccctta attgtacgca ccgctaaaac gcgttcagcg
     1021 cgatcacggc agcagacagg taaaaatggc aacaaaccac ccgaaaaact gccgcaatcg
     1081 cgcctgataa attttaaccg tatgaatacc tatgcaacca gagggtacag gccacattac
     1141 ccccacttaa tccactgaag ctgccatttt tcatggtttc accatcccag cgaagggcca
     1201 tccagcgtgc gttcctgtat ttccggacta taaaaggatc taggtgaaga tcctttttga
     1261 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1321 agaaaagatc aaaggatctt cttgaaatcc tttttttctg cgcgtaatct gctgcttgca
     1381 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1441 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     1501 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1561 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1621 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1681 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1741 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1801 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1861 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1921 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1981 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2041 tgagtgagct gataccgggt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     2101 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagga tccttatcgc
     2161 tatcgattca cacaaaaaac caacacacag atgtaatgaa aataaagata ttttattgcg
     2221 gccatcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     2281 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     2341 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     2401 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     2461 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     2521 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     2581 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     2641 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     2701 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     2761 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     2821 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     2881 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     2941 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     3001 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     3061 atcttgttca atcatgatct gcctcctggg tttagttcct caccttgtcg tattatacta
     3121 tgccgatata ctatgccgat gattaattgt caacagttaa ttaagctttt tgcaaaagcc
     3181 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc
     3241 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     3301 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     3361 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     3421 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     3481 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca
     3541 agttcctctt tcagaggtta tttcaggcca tggtgctgcg cagatccgcg tatgcggtgt
     3601 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     3661 ttttgttaaa attcgatttt accacatttg tagaggtttt acttgcttaa aaaaacctcc
     3721 cacatctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3781 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3841 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3901 gctcgaagcg gccgccccaa ggggttatgc tagttattgc tcagcggtgg cagcagccaa
     3961 ctcagcttcc tttcgggctt tgttagcagc cggatcagct tgcatgcctg caggtcgact
     4021 ctagaggatc cccgggtacc gagcccgaat tcgtttaaac gttcaccacg tcatagtagt
     4081 aattgcgcag ccgcagttcc actgctgcga atccgggcct gttttccaca tcgtatgtcc
     4141 agcacagatt catgagatcg tacatctctc ttggacaccc tgcagggcac cccatccgct
     4201 ctcctttctc taacatagcg gtgacttcac ttcctttcat ccctcgatat ggcttctgcc
     4261 cataggagaa tgcttcccac atcaacactc caaagctcca gacatcgctt ttgctggaga
     4321 acttgtagta gttgatgcat tccggagcgt accacttgac aggccacttt ccatgggtct
     4381 gggccttgta gtagttttca tcagcacgca gtgctttgga aagtccgaaa tcactgatct
     4441 tggcgtaatg ttgggtaact agcaacacat ttcttgcagc cagatctctg tgcacaaaat
     4501 tgctctcctc caagtacttc atgcccatgg aaacctgatg aaccagttct atgatgttct
     4561 tatccttgac atgtctgttc tgctgcaaat acttattgag gggaccaagt tctgccatct
     4621 ccataaccag catccaggac tcggcctcgc atatcccgat catccgcacg atgtacgggt
     4681 tgtccagctg ctgcatgaca tttgcttctg ctaataactc atctttaaga gcggggtcat
     4741 tggcctcgtt tttcagtatt ttcacagcca cggttttcac aacttttttc atttggtagt
     4801 agcccttttt cacagttcca aaattaccag agcccagttc tttgtcttcc agcgtcagca
     4861 gctttcggtc caggtaaacc tccttgggcc tgatctcctc ggggtccgcg taggggctct
     4921 cgtacacctc tgtgtccatg ggtagggctt ctctctgggg gcctttgtct gcagcccagg
     4981 gtgcaagttc tggctcatac ggattgaatg acacagtact ctcttgccgg ttcccttggg
     5041 caggggagga ctttctgtgg ccaggctttg ggaaggagta tgatttgatt cttgagatta
     5101 ttccacccgc tgaccaagtc gcaggatggg aacctggaag ttgtggacgg cctccaaaat
     5161 taacatttcc ctgtgtgccg attttttgac atgggacagt aagaactctt aacaaaccat
     5221 ctgctttata agaataatgc tcgactagct gccagagcgt gtcgaacttc tttccctcgg
     5281 ggatggagag cttccctgtc ttgtctttgt cgatgcgata gtgcagcacc ttcccttcgt
     5341 gcagcaggca cagggcgtag gagccgttgt tgtctctggc tcggatcagg aactttccat
     5401 ttgtctttga tcctatcagg acaatttgct cagattcttc ccgagagatt tttccatgga
     5461 accaaggcat tttttcatgg gctgtggtag cgatcagctt ctccagctga ggcttctgac
     5521 tgatgatggc ctgctccaga gcctgaccct gcaggttcca tgtctgcttc acatattccc
     5581 tgatgaggtt ttccttcaaa tcctcaaagg gcccagtctt gggctgcacc ccttggggcc
     5641 ggttgaaggg cttcttgagg aggcagacca ggccatcaga ctcctgggag tggtagtggc
     5701 agaggtcggc ggggctggca tgggtcctgc caccggcgat ggcgtaggtg ccattcagct
     5761 cccgctcgat ggtgtagtgg tgtgccttcc tcccgtgggc cacggacagg gcgaagccac
     5821 ccaggtagtt gcggctctgg cgcagcaaat aaagcccatc actcatgccc ccctggacca
     5881 ggtaatcttc tgcctcctcc cgggtgatgt tgccgaaaaa gaagggcagg tggttggcgc
     5941 tgtcagccat gccgctgctg gccatggcga tcgcgccgct cgagcccgcc agaatgcgtt
     6001 cgcacagccg ccagccggtc actccgttga tggttactcg gaacagcagg gagccgtcgg
     6061 ggttgatcag gcgctcgtcg ataattttgt tgccgttcca cagggtccct gttacagtga
     6121 tctttttgcc gtcgaacacg gcgatgcctt catacggccg tccgaaatag tcgatcatgt
     6181 tcggcgtaac cccgtcgatt accagtgtgc catagtgcag gatcacctta aagtgatgat
     6241 catccacagg gtacaccacc ttaaaaattt tttcgatctg gcccatttgg tcgccgctca
     6301 gaccttcata cgggatgatg acatggatgt cgatcttcag cccattttca ccgctcagga
     6361 caatcctttg gatcggagtt acggacaccc cgagattctg aaacaaactg gacacacctc
     6421 cctgttcaag gacttggtcc aggttgtagc cggctgtctg tcgccagtcc ccaacgaaat
     6481 cttcgagtgt gaagaccatg gtgagcgcta gccctatagt gagtcgtatt aatactctag
     6541 ccttaagagc tgtaattgaa ctgggagtgg acacctgtgg agagaaaggc aaagtggatg
     6601 tcagtaagac caataggtgc ctatcagaaa cgcaagagtc ttctctgtct cgacaagccc
     6661 agtttctatt ggtc
//
