LOCUS       sequence-478464-        5588 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HDAC1-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 5588)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5588)
  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            "Unpublished"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: HDAC1-HiBiT Vector
FEATURES             Location/Qualifiers
     source          1..5588
                     /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(2667..2685)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2722..2843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2733..2752)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2787..2806
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3088..3445
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3088..3279
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3111..3131)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3296..3431
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3460..3507
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3539..4333
                     /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(3593..3612)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4203..4222
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4397..4445
                     /note="synthetic polyadenylation signal"
     rep_origin      4602..5147
                     /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     5048..5067
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5264..5281
                     /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 cagacgcagg
     1081 gcacccggag gaaagtctgt tactactacg acggggatgt tggaaattac tattatggac
     1141 aaggccaccc aatgaagcct caccgaatcc gcatgactca taatttgctg ctcaactatg
     1201 gtctctaccg aaaaatggaa atctatcgcc ctcacaaagc caatgctgag gagatgacca
     1261 agtaccacag cgatgactac attaaattct tgcgctccat ccgtccagat aacatgtcgg
     1321 agtacagcaa gcagatgcag agattcaacg ttggtgagga ctgtccagta ttcgatggcc
     1381 tgtttgagtt ctgtcagttg tctactggtg gttctgtggc aagtgctgtg aaacttaata
     1441 agcagcagac ggacatcgct gtgaattggg ctgggggcct gcaccatgca aagaagtccg
     1501 aggcatctgg cttctgttac gtcaatgata tcgtcttggc catcctggaa ctgctaaagt
     1561 atcaccagag ggtgctgtac attgacattg atattcacca tggtgacggc gtggaagagg
     1621 ccttctacac cacggaccgg gtcatgactg tgtcctttca taagtatgga gagtacttcc
     1681 caggaactgg ggacctacgg gatatcgggg ctggcaaagg caagtattat gctgttaact
     1741 acccgctccg agacgggatt gatgacgagt cctatgaggc cattttcaag ccggtcatgt
     1801 ccaaagtaat ggagatgttc cagcctagtg cggtggtctt acagtgtggc tcagactccc
     1861 tatctgggga tcggttaggt tgcttcaatc taactatcaa aggacacgcc aagtgtgtgg
     1921 aatttgtcaa gagctttaac ctgcctatgc tgatgctggg aggcggtggt tacaccattc
     1981 gtaacgttgc ccggtgctgg acatatgaga cagctgtggc cctggatacg gagatcccta
     2041 atgagcttcc atacaatgac tactttgaat actttggacc agatttcaag ctccacatca
     2101 gtccttccaa tatgactaac cagaacacga atgagtacct ggagaagatc aaacagcgac
     2161 tgtttgagaa ccttagaatg ctgccgcacg cacctggggt ccaaatgcag gcgattcctg
     2221 aggacgccat ccctgaggag agtggcgatg aggacgaaga cgaccctgac aagcgcatct
     2281 cgatctgctc ctctgacaaa cgaattgcct gtgaggaaga gttctccgat tctgaagagg
     2341 agggagaggg gggccgcaag aactcttcca acttcaaaaa agccaagaga gtcaaaacag
     2401 aggatgaaaa agagaaagac ccagaggaga agaaagaagt caccgaagag gagaaaacca
     2461 aggaggagaa gccagaagcc aaaggggtca aggaggaggt caagttggcc gtttctcaag
     2521 gcagttcagg tggctcgagc ggtgtgagcg gctggcggct gttcaagaag attagctaag
     2581 gccgcgactc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg
     2641 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     2701 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2761 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2821 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2881 agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg
     2941 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     3001 tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc
     3061 tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg
     3121 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3181 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3241 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3301 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3361 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3421 ttttggaggc ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca
     3481 tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat
     3541 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3601 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3661 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3721 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3781 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3841 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3901 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3961 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4021 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4081 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4141 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4201 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4261 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4321 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4381 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4441 gtgtgaatcg atagcgataa ggatccgctc atgcgatgat aagctgtcaa acatgagaat
     4501 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
     4561 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
     4621 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     4681 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     4741 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     4801 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     4861 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     4921 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     4981 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     5041 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     5101 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     5161 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     5221 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     5281 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     5341 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     5401 cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcatgag
     5461 attacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc
     5521 cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat
     5581 cgagatct
//
