LOCUS       sequence-473383-        5748 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express CLK3-HiBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5748)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5748)
  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: CLK3-HiBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5748
                     /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"
     primer_bind     complement(2691..2709)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2746..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2757..2776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2811..2830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3112..3469
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3112..3303
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3135..3155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3320..3455
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3484..3531
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3563..4357
                     /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(3617..3636)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4227..4246
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4421..4469
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4708..5296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4788..4807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5412..5695
                     /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 cgccatgcac cactgtaaac
     1081 ggtaccgatc accggaacca gatccctacc tcagctatcg atggaagcgc cgccggagtt
     1141 attctcgaga acatgaagga cgccttcgct atccgtcccg cagagaaccg cccccaaggc
     1201 gaagtcgatc acgcagtcat gaccgccttc cctatcaacg gaggtatcgg gagcgaagag
     1261 acagcgatac atatcggtgt gaggaacggt ctcctagctt cggcgaagat tactacggtc
     1321 ccagtcgaag ccgccatcga cgcaggtcca gggaaagggg accgtatcga acaagaaagc
     1381 acgctcatca ctgccataaa cgacggacta ggtcttgtag cagtgctagc tcacgaagtc
     1441 aacagtcatc taagcgcagc tcaagatctg tagaggacga caaagaagga catcttgtat
     1501 gccgaatcgg tgattggctt caggaacggt atgagattgt gggaaatttg ggagaaggaa
     1561 catttgggaa ggttgtggag tgtttggacc acgctcgcgg taaaagtcaa gtagctctca
     1621 aaatcatccg caatgtaggg aagtatcgcg aggcggcgcg gctcgaaatc aacgtgctta
     1681 agaagattaa ggagaaggac aaagagaaca aatttctttg cgttctcatg agtgactggt
     1741 ttaactttca tggccacatg tgcattgcct ttgagctcct gggcaagaac acctttgagt
     1801 tcctgaagga gaataacttc cagccttacc ccctaccaca tgtccggcac atggcctacc
     1861 agctctgcca cgcccttaga tttctgcatg agaatcagct gacccataca gacttgaaac
     1921 cagagaacat cctgtttgtg aattctgagt ttgaaaccct ctacaatgag cacaagagct
     1981 gtgaggagaa gtcagtgaag aacaccagca tccgagtggc tgactttggc agtgccacat
     2041 ttgaccatga gcaccacacc accattgtgg ccacccgtca ctatcgcccg cctgaggtga
     2101 tccttgagct gggctgggca cagccctgtg acgtctggag cattggctgc attctctttg
     2161 agtactaccg gggcttcaca ctcttccaga cccacgaaaa ccgagagcac ctggtgatga
     2221 tggagaagat cctagggccc atcccatcac acatgatcca ccgtaccagg aagcagaaat
     2281 atttctacaa agggggccta gtttgggatg agaacagctc tgacggccgg tatgtgaagg
     2341 agaactgcaa acctctgaag agttacatgc tccaagactc cctggagcac gtgcagctgt
     2401 ttgacctgat gaggaggatg ttagaatttg accctgccca gcgcatcaca ctggccgagg
     2461 ccctgctgca ccccttcttt gctggcctga cccctgagga gcggtccttc cacaccagcc
     2521 gcaacccaag cagagtttct caaggcagtt caggtggctc gagcggtgtg agcggctggc
     2581 ggctgttcaa gaagattagc taaggccgcg actctagagt cgacctgcag gcatgcaagc
     2641 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2701 ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg
     2761 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2821 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2881 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa
     2941 acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga
     3001 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     3061 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     3121 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3301 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3421 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta
     3481 actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     3541 aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc
     3601 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3661 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3721 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3781 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3841 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3901 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3961 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4021 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4081 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4141 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4201 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4261 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4321 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4381 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     4441 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct
     4501 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     4561 ctgcggactg gctttctacc cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4621 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4681 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5161 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5281 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5401 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     5461 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     5521 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     5581 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     5641 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     5701 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
