LOCUS       sequence-471860-        5067 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express FKBP-12-NanoLuc(R) 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 5067)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5067)
  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: FKBP-12-NanoLuc Vector
FEATURES             Location/Qualifiers
     source          1..5067
                     /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             join(1065..1067,1068..1388)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             1410..1922
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2010..2028)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2065..2186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2076..2095)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2130..2149
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2431..2788
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2431..2622
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2454..2474)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2639..2774
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2803..2850
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2882..3676
                     /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(2936..2955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3546..3565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3740..3788
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4027..4615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4107..4126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4731..5014
                     /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 ggctagcgat cgccatggga gtgcaggtgg
     1081 aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
     1141 actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc
     1201 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
     1261 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
     1321 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
     1381 aactggaagt ttctctcggc tcgagcggcg tcttcacact cgaagatttc gttggggact
     1441 ggcgacagac agccggctac aacctggacc aagtccttga acagggaggt gtgtccagtt
     1501 tgtttcagaa tctcggggtg tccgtaactc cgatccaaag gattgtcctg agcggtgaaa
     1561 atgggctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc ggcgaccaaa
     1621 tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat cactttaagg
     1681 tgatcctgca ctatggcaca ctggtaatcg acggggttac gccgaacatg atcgactatt
     1741 tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact gtaacaggga
     1801 ccctgtggaa cggcaacaaa attatcgacg agcgcctgat caaccccgac ggctccctgc
     1861 tgttccgagt aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc attctggcgt
     1921 aaggccgcga ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc
     1981 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2041 ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     2101 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2161 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2221 ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa
     2281 tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     2341 atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac
     2401 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     2461 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2521 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     2581 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     2641 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2701 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2761 cttttttgga ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg
     2821 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat
     2881 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2941 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3001 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3061 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3121 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3181 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3241 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3301 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3361 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3421 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3481 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3541 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3601 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3661 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3721 ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     3781 tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc ttgtccagat
     3841 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctaccc
     3901 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3961 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4441 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     4621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac
     4741 aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc
     4801 agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt
     4861 taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg
     4921 tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt
     4981 aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga
     5041 tgcgtccggc gtagaggatc gagatct
//
