LOCUS       sequence-474688-        5725 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express NanoLuc(R)-PP1Cg 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 5725)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5725)
  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: NanoLuc-PP1Cg Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5725
                     /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(2681..2699)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2736..2857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2747..2766)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2801..2820
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3102..3459
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3102..3293
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3125..3145)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3310..3445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3540..4334
                     /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(3594..3613)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4204..4223
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4398..4446
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4685..5273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4765..4784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5389..5672
                     /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 ggcggattta gataaactca
     1621 acatcgacag cattatccaa cggctgctgg aagtgagagg gtccaagcct ggtaagaatg
     1681 tccagcttca ggagaatgaa atcagaggac tgtgcttaaa gtctcgtgaa atctttctca
     1741 gtcagcctat cctactagaa cttgaagcac cactcaaaat atgtggtgac atccatggac
     1801 aatactatga tttgctgcga ctttttgagt acggtggttt cccaccagaa agcaactacc
     1861 tgtttcttgg ggactatgtg gacaggggaa agcagtcatt ggagacgatc tgcctcttac
     1921 tggcctacaa aataaaatat cctgagaatt tttttcttct cagagggaac catgaatgtg
     1981 ccagcatcaa cagaatttat ggattttatg atgaatgtaa aagaagatac aacattaaac
     2041 tatggaaaac tttcacagac tgttttaact gtttaccgat agcagccatc gtggatgaga
     2101 agatattctg ctgtcatgga ggtttatcac cagatcttca atctatggag cagattcggc
     2161 gaattatgcg accaactgat gtaccagatc aaggtcttct ttgtgatctt ttgtggtctg
     2221 accccgataa agatgtctta ggctggggtg aaaatgacag aggagtgtcc ttcacatttg
     2281 gtgcagaagt ggttgcaaaa tttctccata agcatgattt ggatcttata tgtagagccc
     2341 atcaggtggt tgaagatgga tatgaatttt ttgcaaagag gcagttggtc actctgtttt
     2401 ctgcgcccaa ttattgcgga gagtttgaca atgcaggtgc catgatgagt gtggatgaaa
     2461 cactaatgtg ttcttttcag attttaaagc ctgcagagaa aaagaagcca aatgccacga
     2521 gacctgtaac gcctccaagg ggtatgatca caaagcaagc aaagaaagtt taaacgaatt
     2581 cgggctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc tgatccggct
     2641 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     2701 taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2761 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2821 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2881 tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa
     2941 atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     3001 tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg
     3061 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     3121 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3361 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3421 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta actgttgaca
     3481 attaatcatc ggcatagtat aatacgacaa ggcgaggaac taaacccagg aggcagatca
     3541 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3601 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3661 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3721 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3781 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3841 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3901 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3961 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4021 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4081 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4141 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4201 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4261 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4321 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4381 gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     4441 tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt gtccagatag
     4501 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacccgg
     4561 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4621 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4681 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4741 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4801 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4861 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4921 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4981 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5041 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5101 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5161 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5221 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     5281 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5341 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaaatacag
     5401 gaacgcacgc tggatggccc ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag
     5461 tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta
     5521 aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc
     5581 tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa
     5641 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
     5701 cgtccggcgt agaggatcga gatct
//
