LOCUS       sequence-478466-        5258 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express CDK9-HiBiT 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 5258)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5258)
  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            "Unpublished"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: CDK9-HiBiT Vector
FEATURES             Location/Qualifiers
     source          1..5258
                     /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(2337..2355)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2392..2513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2403..2422)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2457..2476
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2758..3115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2758..2949
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2781..2801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2966..3101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3130..3177
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3209..4003
                     /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(3263..3282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3873..3892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4067..4115
                     /note="synthetic polyadenylation signal"
     rep_origin      4272..4817
                     /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     4718..4737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4934..4951
                     /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 cgccatggcg aagcagtacg
     1081 actcggtgga gtgccctttt tgtgatgaag tttccaaata cgagaagctc gccaagatcg
     1141 gccaaggcac cttcggggag gtgttcaagg ccaggcaccg caagaccggc cagaaggtgg
     1201 ctctgaagaa ggtgctgatg gaaaacgaga aggaggggtt ccccattaca gccttgcggg
     1261 agatcaagat ccttcagctt ctaaaacacg agaatgtggt caacttgatt gagatttgtc
     1321 gaaccaaagc ttccccctat aaccgctgca agggtagtat atacctggtg ttcgacttct
     1381 gcgagcatga ccttgctggg ctgttgagca atgttttggt caagttcacg ctgtctgaga
     1441 tcaagagggt gatgcagatg ctgcttaacg gcctctacta catccacaga aacaagatcc
     1501 tgcataggga catgaaggct gctaatgtgc ttatcactcg tgatggggtc ctgaagctgg
     1561 cagactttgg gctggcccgg gccttcagcc tggccaagaa cagccagccc aaccgctaca
     1621 ccaaccgtgt ggtgacactc tggtaccggc ccccggagct gttgctcggg gagcgggact
     1681 acggcccccc cattgacctg tggggtgctg ggtgcatcat ggcagagatg tggacccgca
     1741 gccccatcat gcagggcaac acggagcagc accaactcgc cctcatcagt cagctctgcg
     1801 gctccatcac ccctgaggtg tggccaaacg tggacaacta tgagctgtac gaaaagctgg
     1861 agctggtcaa gggccagaag cggaaggtga aggacaggct gaaggcctat gtgcgtgacc
     1921 catacgcact ggacctcatc gacaagctgc tggtgctgga ccctgcccag cgcatcgaca
     1981 gcgatgacgc cctcaaccac gacttcttct ggtccgaccc catgccctcc gacctcaagg
     2041 gcatgctctc cacccacctg acgtccatgt tcgagtactt ggcaccaccg cgccggaagg
     2101 gcagccagat cacccagcag tccaccaacc agagtcgcaa tcccgccacc accaaccaga
     2161 cggagtttga gcgcgtcttc gtttctcaag gcagttcagg tggctcgagc ggtgtgagcg
     2221 gctggcggct gttcaagaag attagctaag gccgcgactc tagagtcgac ctgcaggcat
     2281 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2341 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     2401 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2461 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2521 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     2581 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     2641 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     2701 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     2761 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2821 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2881 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2941 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3001 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3061 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3121 taattaactg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3181 gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc
     3241 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3301 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3361 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3421 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3481 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3541 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3601 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3721 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4021 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     4081 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgctc
     4141 atgcgatgat aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca
     4201 ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata
     4261 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     4321 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     4381 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     4441 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     4501 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     4561 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     4621 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     4681 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     4741 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     4801 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     4861 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     4921 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     4981 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     5041 tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc
     5101 ttttgccgtt acgcaccacc ccgtcatgag attacgcgtt ttagcggtgc gtacaattaa
     5161 gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac
     5221 tccggccacg atgcgtccgg cgtagaggat cgagatct
//
