LOCUS       sequence-474686-        6504 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express NanoLuc(R)-SHOC2 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 6504)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay, NanoLuc-SHOC2 Vector
FEATURES             Location/Qualifiers
     source          1..6504
                     /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(3458..3476)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3513..3634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3524..3543)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3578..3597
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3879..4236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3879..4070
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3902..3922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4087..4222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4251..4298
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4330..5124
                     /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(4384..4403)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4994..5013
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5188..5236
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5464..6052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5544..5563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6168..6451
                     /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 gagtagtagt ttaggaaaag
     1621 aaaaagactc taaagaaaaa gatcccaaag taccatcagc caaggaaaga gaaaaggagg
     1681 caaaagcctc tggaggtttt gggaaagaga gcaaagaaaa agaacctaag accaaaggga
     1741 aagatgccaa agatggaaag aaggactcca gtgctgccca accaggggtg gcattttcag
     1801 ttgacaatac gatcaaacgg ccaaacccag cacctgggac tagaaaaaaa tccagcaatg
     1861 cagaggtgat taaagagctc aacaaatgcc gggaagagaa ttcaatgcgt ttggacttat
     1921 ccaagagatc tatacacata ttgccatcat caatcaaaga gttgactcaa ttaacagaac
     1981 tttatttata cagtaacaaa ttgcagtccc tcccagcaga ggtgggatgt ttagtaaatc
     2041 tcatgacact ggctctaagt gaaaattcac ttaccagttt gcctgactct cttgataact
     2101 tgaagaagct gcggatgctt gatttacggc ataataaact gagagaaatt ccttcagtgg
     2161 tgtataggct ggattctctc accactcttt accttcgctt taatcgtata actactgtgg
     2221 aaaaggacat caaaaacttg tcaaaactca gcatgcttag cattcgagag aacaaaatta
     2281 aacaactacc tgctgaaatt ggtgaattat gtaacctcat tacgctggat gtagctcaca
     2341 atcaacttga acaccttcca aaggagattg gaaactgtac acagataacc aaccttgact
     2401 tgcagcacaa tgaactgcta gacctcccag atactatagg aaacctgtcc agtttaagtc
     2461 gtcttggtct gagatataac agactgtcag caatacccag atcattagca aaatgcagtg
     2521 cacttgaaga attaaattta gagaacaata acatttctac tttaccagag agtcttttat
     2581 caagtcttgt gaaactgaat agtttgacct tagctagaaa ttgcttccag ttgtatccag
     2641 tgggtggtcc atctcagttt tctaccatct attccctcaa catggaacac aatcgaatca
     2701 acaaaattcc atttggaatt ttctccagag caaaagtatt aagtaagctg aatatgaagg
     2761 acaatcagtt aacatcactt cccttggatt ttggaacttg gaccagtatg gtagaattga
     2821 atttagccac taatcagctc acaaagatcc ctgaggatgt gtctggtctc gtttctcttg
     2881 aggttcttat cttatcaaac aatcttctaa agaagcttcc ccatggtctt ggaaacctta
     2941 ggaagttaag agagttggat ctagaagaga acaaattgga atccttgcca aatgaaattg
     3001 catatcttaa ggatttacag aaattagtct tgacaaacaa ccagttgacc actcttccca
     3061 gaggcattgg tcaccttact aatctcacac atctgggcct tggagagaac ctacttactc
     3121 accttcctga agaaattggt acactggaga acctagaaga actgtatttg aatgacaacc
     3181 ccaacctgca tagccttccc tttgagctgg cactctgcag caagctttca atcatgagta
     3241 ttgagaactg tccactcagt caccttccac ctcagattgt tgctgggggg ccttctttca
     3301 tcattcagtt cttaaagatg cagggtccat atcgtgccat ggtcgtttaa acgaattcgg
     3361 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagctga tccggctgct
     3421 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     3481 ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3541 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3601 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3661 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     3721 tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     3781 ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct
     3841 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     3901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4141 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact gttgacaatt
     4261 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     4321 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4381 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4441 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4501 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4561 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4621 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4681 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4741 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4801 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4861 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4921 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4981 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5041 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5101 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5161 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     5221 gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt
     5281 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     5341 ttctacccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5401 aacgcaggaa agaacatgtg agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5461 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5521 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5581 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5641 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5701 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5761 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5821 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5881 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     5941 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6001 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     6061 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6121 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg
     6181 aacgcacgct ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt
     6241 ggattaagtg ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa
     6301 aatttatcag gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct
     6361 gctgccgtga tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa
     6421 tccacttact gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc
     6481 gtccggcgta gaggatcgag atct
//
