LOCUS       sequence-479902-        5196 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express CDK2-SmBiT 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 5196)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5196)
  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            "Unpublished"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: CDK2-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2259..2380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2270..2289)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2495..2528)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2538..2585
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2617..3411
                     /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(2671..2690)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3281..3300
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3649..3668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4283..4331
                     /note="synthetic polyadenylation signal"
     CDS             complement(4355..4753)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4750..4759
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4784..5141)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4798..4933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4950..5141
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5098..5118
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgatcgccat ggagaacttc
     1201 caaaaggtgg aaaagatcgg agagggcacg tacggagttg tgtacaaagc cagaaacaag
     1261 ttgacgggag aggtggtggc gcttaagaaa atccgcctgg acactgagac tgagggtgtg
     1321 cccagtactg ccatccgaga gatctctctg cttaaggagc ttaaccatcc taatattgtc
     1381 aagctgctgg atgtcattca cacagaaaat aaactctacc tggtttttga atttctgcac
     1441 caagatctca agaaattcat ggatgcctct gctctcactg gcattcctct tcccctcatc
     1501 aagagctatc tgttccagct gctccagggc ctagctttct gccattctca tcgggtcctc
     1561 caccgagacc ttaaacctca gaatctgctt attaacacag agggggccat caagctagca
     1621 gactttggac tagccagagc ttttggagtc cctgttcgta cttacaccca tgaggtggtg
     1681 accctgtggt accgagctcc tgaaatcctc ctgggctgca aatattattc cacagctgtg
     1741 gacatctgga gcctgggctg catctttgct gagatggtga ctcgccgggc cctattccct
     1801 ggagattctg agattgacca gctcttccgg atctttcgga ctctggggac cccagatgag
     1861 gtggtgtggc caggagttac ttctatgcct gattacaagc caagtttccc caagtgggcc
     1921 cggcaagatt ttagtaaagt tgtacctccc ctggatgaag atggacggag cttgttatcg
     1981 caaatgctgc actacgaccc taacaagcgg atttcggcca aggcagccct ggctcaccct
     2041 ttcttccagg atgtgaccaa gccagtaccc catcttcgac tcgtttctca aggcagttca
     2101 ggtggtggcg ggagcggagg tggaggctcg agcggtgtga ccggctaccg gctgttcgag
     2161 gagattctgt aaggccgcga ctctagagtc gacctgcagg catgcaagct gatttggctg
     2221 ctgccaccgc tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2281 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2341 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2401 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     2461 caaatgtggt aaaatcttaa ttaagaattc taatataact tcgtatagca tacattatac
     2521 gaagttatgg atccactgtt gacaattaat catcggcata gtatatcggc atagtataat
     2581 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     2641 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     2701 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     2761 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     2821 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     2881 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     2941 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3001 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3061 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3121 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3181 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     3241 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     3301 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     3361 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg
     3421 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4261 tccttttata gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     4321 gatattttat tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg
     4381 gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt
     4441 tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc
     4501 ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag
     4561 cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac
     4621 cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga
     4681 tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca
     4741 aaggcttggc catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc
     4801 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     4861 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     4921 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     4981 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     5041 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     5101 ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc
     5161 ctctttcaga ggttatttca ggccatggtg ctgcgc
//
