LOCUS       sequence-486830-        5513 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Express HiBiT-CHRM1 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5513)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-CHRM1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        272..475
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     426..446
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          611..743
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     726..745
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     787..806
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        787..805
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2210..2228)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2265..2386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2276..2295)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2330..2349
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2631..2988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2631..2822
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2654..2674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2839..2974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3003..3050
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3082..3876
                     /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(3136..3155)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3746..3765
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3940..3988
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4227..4815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4307..4326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4931..5214
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        join(5405..5513,1..271)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
       61 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      121 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa
      181 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta
      241 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg
      301 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
      361 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc
      421 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
      481 agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt aaattgctaa
      541 cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg
      601 cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
      661 cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat
      721 ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta
      781 gagtattaat acgactcact atagggctag cgctcaccat gaactccttc tccacaagcg
      841 ccttcggtcc agttgccttc tccctgggcc tgctcctggt gttgcctgct gccttccctg
      901 ccccagtctc cgtgagcggc tggcggctgt tcaagaagat tagcgggagc tccggtggct
      961 cgagcggtgc gatcgccatg accgaggtgc tgtggccggc tgtccccaac gggacggacg
     1021 ctgccttcct ggccggtccg ggttcgtcct gggggaacag cacggtcgcc tccactgccg
     1081 ccgtctcctc gtcgttcaaa tgcgccttga ccaagacggg cttccagttt tactacctgc
     1141 cggctgtcta catcttggta ttcatcatcg gcttcctggg caacagcgtg gccatctgga
     1201 tgttcgtctt ccacatgaag ccctggagcg gcatctccgt gtacatgttc aatttggctc
     1261 tggccgactt cttgtacgtg ctgactctgc cagccctgat cttctactac ttcaataaaa
     1321 cagactggat cttcggggat gccatgtgta aactgcagag gttcatcttt catgtgaacc
     1381 tctatggcag catcttgttt ctgacatgca tcagtgccca ccggtacagc ggtgtggtgt
     1441 accccctcaa gtccctgggc cggctcaaaa agaagaatgc gatctgtatc agcgtgctgg
     1501 tgtggctcat tgtggtggtg gcgatctccc ccatcctctt ctactcaggt accggggtcc
     1561 gcaaaaacaa aaccatcacc tgttacgaca ccacctcaga cgagtacctg cgaagttatt
     1621 tcatctacag catgtgcacg accgtggcca tgttctgtgt ccccttggtg ctgattctgg
     1681 gctgttacgg attaattgtg agagctttga tttacaaaga tctggacaac tctcctctga
     1741 ggagaaaatc gatttacctg gtaatcattg tactgactgt ttttgctgtg tcttacatcc
     1801 ctttccatgt gatgaaaacg atgaacttga gggcccggct tgattttcag accccagcaa
     1861 tgtgtgcttt caatgacagg gtttatgcca cgtatcaggt gacaagaggt ctagcaagtc
     1921 tcaacagttg tgtggacccc attctctatt tcttggcggg agatactttc agaaggagac
     1981 tctcccgagc cacaaggaaa gcttctagaa gaagtgaggc aaatttgcaa tccaagagtg
     2041 aagacatgac cctcaatatt ttacctgagt tcaagcagaa tggagataca agcctggttt
     2101 aaacgaattc gggctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
     2161 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     2221 taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga
     2281 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2341 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2401 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     2461 cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat
     2521 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     2581 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     2641 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2701 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2761 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2821 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2881 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2941 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaattaa
     3001 ctgttgacaa ttaatcatcg gcatagtatg tcggcatagt ataatacgac aaggtgagga
     3061 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3121 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3181 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3241 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3301 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3361 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3421 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3481 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3541 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3601 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3661 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3721 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3781 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3841 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3901 ccgaccaagc gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca
     3961 ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt
     4021 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     4081 tgcggactgg ctttctaccc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4141 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4201 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4261 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4321 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4381 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4441 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4501 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4561 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4621 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4681 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4801 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4861 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4921 cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa
     4981 ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt
     5041 tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg
     5101 tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg
     5161 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     5221 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatcttca atattggcca
     5281 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     5341 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     5401 tgttggcatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     5461 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tgg
//
