LOCUS       sequence-479803-        5849 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express STK17A-NanoLuc 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 5849)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: STK17A-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5849
                     /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             2328..2840
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2928..2946)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2983..3104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2994..3013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3048..3067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3349..3706
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3349..3540
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3372..3392)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3557..3692
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3721..3768
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3800..4594
                     /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(3854..3873)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4464..4483
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4658..4706
                     /note="synthetic polyadenylation signal"
     rep_origin      4863..5408
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5309..5328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5525..5542
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 ggctagcgat cgccatgatc cctttggaga
     1081 agccaggcag cggcggctcc tccccaggcg ccacctcagg ctcgggccgg gcaggccggg
     1141 gtctgagcgg gccgtgccgg ccgccgccgc cgccccaggc ccgcgggctg ttgacagaga
     1201 tacgcgccgt ggtgcgcacc gagcccttcc aggacggcta cagcctgtgc ccgggccggg
     1261 agctgggcag ggggaaattt gcagtggtga gaaaatgtat aaagaaagat tctgggaaag
     1321 aatttgctgc aaagttcatg agaaaaagaa gaaaaggcca agattgtcgg atggaaataa
     1381 ttcatgagat tgctgtactt gaactagcac aagacaatcc ttgggtcatt aatttacatg
     1441 aagtttatga gactgcatca gaaatgatct tagttctgga atatgctgct gggggtgaaa
     1501 tctttgacca gtgtgttgca gacagagaag aagcctttaa agaaaaagat gttcaaagac
     1561 ttatgcgaca gattttagaa ggtgttcact ttttacacac tcgtgatgta gttcatcttg
     1621 atttgaagcc tcagaatatt ctgttgacaa gtgaatctcc attgggtgac attaagattg
     1681 ttgattttgg cctttcaaga atattgaaga acagtgaaga gctccgagaa attatgggta
     1741 cccctgaata tgtggctcct gaaattctta gttatgatcc tataagcatg gcaacagata
     1801 tgtggagcat tggagtgtta acatatgtca tgcttacagg aatatcacct ttcttaggca
     1861 atgataaaca agaaacattc ttaaacatct cacagatgaa tttaagttat tctgaggaag
     1921 aatttgatgt tttgtctgag tcggctgttg atttcatcag gacactttta gttaagaaac
     1981 ctgaagatcg agccactgct gaagaatgtc taaagcaccc ctggttgaca cagagcagta
     2041 ttcaagagcc ttctttcagg atggaaaagg cactagaaga agcaaatgcc ctccaagaag
     2101 gtcattctgt gcctgaaatt aattcggata ccgacaaatc agaaaccgag gaatccattg
     2161 taaccgaaga gttaattgta gttacttcat atactctagg acaatgcaga cagtctgaaa
     2221 aagagaaaat ggagcaaaag gccatttcca aacgatttaa atttgaggaa cctttgctac
     2281 aagaaattcc aggagaattt atctacgttt ctctcggctc gagcggcgtc ttcacactcg
     2341 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     2401 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     2461 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     2521 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     2581 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     2641 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     2701 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     2761 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     2821 gcgaacgcat tctggcgtaa ggccgcgact ctagagtcga cctgcaggca tgcaagctga
     2881 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     2941 actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt
     3001 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3061 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3121 ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc
     3181 tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     3241 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     3301 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     3361 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3421 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3541 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3601 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3661 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact
     3721 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     3781 taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3841 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3901 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3961 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4021 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4081 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4141 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4201 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4261 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4321 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4381 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4441 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4501 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4561 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4621 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     4681 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatccgct catgcgatga
     4741 taagctgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca
     4801 tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct
     4861 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     4921 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     4981 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     5041 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     5101 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     5161 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     5221 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     5281 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     5341 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     5401 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     5461 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     5521 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     5581 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     5641 ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt
     5701 tacgcaccac cccgtcatga gattacgcgt tttagcggtg cgtacaatta agggattatg
     5761 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     5821 gatgcgtccg gcgtagagga tcgagatct
//
