LOCUS       sequence-478463-        5807 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-HTR2A 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 5807)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5807)
  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            "Unpublished"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: HiBiT-HTR2A Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5807
                     /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(2750..2768)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2805..2926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2816..2835)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2870..2889
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3171..3528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3171..3362
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3194..3214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3379..3514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3543..3590
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3622..4416
                     /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(3676..3695)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4286..4305
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4480..4528
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4767..5355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4847..4866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5471..5754
                     /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 ggctagcgct caccatgaac tccttctcca
     1081 caagcgcctt cggtccagtt gccttctccc tgggcctgct cctggtgttg cctgctgcct
     1141 tccctgcccc agtctccgtg agcggctggc ggctgttcaa gaagattagc gggagctccg
     1201 gtggctcgag cggtgcgatc gccatggata ttctttgtga agaaaatact tctttgagct
     1261 caactacgaa ctccctaatg caattaaatg atgacaccag gctctacagt aatgacttta
     1321 actctggaga agctaacact tctgatgcat ttaactggac agtcgactct gaaaatcgaa
     1381 ccaacctttc ctgtgaaggg tgcctctcac cgtcgtgtct ctccttactt catctccagg
     1441 aaaaaaactg gtctgcttta ctgacagccg tagtgattat tctaactatt gctggaaaca
     1501 tactcgttat catggcagtg tccctagaga aaaagctgca gaatgccacc aactatttcc
     1561 tgatgtcact tgccatagct gatatgctgc tgggtttcct tgtcatgccc gtgtccatgt
     1621 taaccatcct gtatgggtac cggtggcctc tgccgagcaa gctttgtgca gtctggattt
     1681 acctggacgt gctcttctcc acggcctcca tcatgcacct ctgcgccatc tcgctggacc
     1741 gctacgtcgc catccagaat cccatccacc acagccgctt caactccaga actaaggcat
     1801 ttctgaaaat cattgctgtt tggaccatat cagtaggtat atccatgcca ataccagtct
     1861 ttgggctaca ggacgattcg aaggtcttta aggaggggag ttgcttactc gccgatgata
     1921 actttgtcct gatcggctct tttgtgtcat ttttcattcc cttaaccatc atggtgatca
     1981 cctactttct aactatcaag tcactccaga aagaagctac tttgtgtgta agtgatcttg
     2041 gcacacgggc caaattagct tctttcagct tcctccctca gagttctttg tcttcagaaa
     2101 agctcttcca gcggtcgatc catagggagc cagggtccta cacaggcagg aggactatgc
     2161 agtccatcag caatgagcaa aaggcatgca aggtgctggg catcgtcttc ttcctgtttg
     2221 tggtgatgtg gtgccctttc ttcatcacaa acatcatggc cgtcatctgc aaagagtcct
     2281 gcaatgagga tgtcattggg gccctgctca atgtgtttgt ttggatcggt tatctctctt
     2341 cagcagtcaa cccactagtc tacacactgt tcaacaagac ctataggtca gccttttcac
     2401 ggtatattca gtgtcagtac aaggaaaaca aaaaaccatt gcagttaatt ttagtgaaca
     2461 caataccggc tttggcctac aagtctagcc aacttcaaat gggacaaaaa aagaattcaa
     2521 agcaagatgc caagacaaca gataatgact gctcaatggt tgctctagga aagcagcatt
     2581 ctgaagaggc ttctaaagac aatagcgacg gagtgaatga aaaggtgagc tgtgtggttt
     2641 aaacgaattc gggctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
     2701 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2761 taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga
     2821 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2881 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2941 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     3001 cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat
     3061 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     3121 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     3181 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3361 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaattaa
     3541 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3601 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3661 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3721 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3781 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3841 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3901 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3961 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4021 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4081 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4141 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4201 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4261 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4321 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4381 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4441 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     4501 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt
     4561 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     4621 tgcggactgg ctttctaccc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4681 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5281 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5341 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5401 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5461 cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa
     5521 ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt
     5581 tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg
     5641 tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg
     5701 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     5761 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
