LOCUS       sequence-473538-        5682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CHRM1-SmBiT 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 5682)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5682)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: CHRM1-SmBiT Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(747..1541)
                     /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(858..877)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1468..1487
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1573..1620)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    1630..1663
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1869..1888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3468..3487)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3469..3487)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3529..3548)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3531..3663
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(3799..4002)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3828..3848)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4003..4306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4539..4620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4699..5056
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4699..4890
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4722..4742)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4907..5042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      5081..5090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5087..5485
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5509..5557
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccggttg
      721 ggcgtcgctt ggtcggtcat ttcgaatcag aagaactcgt caagaaggcg atagaaggcg
      781 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
      841 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
      901 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
      961 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     1021 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     1081 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     1141 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1201 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1261 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1321 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     1381 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1441 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     1501 gcggccggag aacctgcgtg caatccatct tgttcaatca tgatctgcct cctgggttta
     1561 gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     1621 agtggatcca taacttcgta taatgtatgc tatacgaagt tatattagaa ttcttaatta
     1681 agattttacc acatttgtag aggttttact tgcttaaaaa aacctcccac atctccccct
     1741 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc
     1921 ttgctcagcg gtggcagcag ccaaatcagc ttgcatgcct gcaggtcgac tctagagtcg
     1981 cggccttaca gaatctcctc gaacagccgg tagccggtca caccgctcga gcctccacct
     2041 ccgctcccgc caccacctga actgccttga gaaacgcatt ggcgggaggg agtgcggtgc
     2101 acggagccag ggcgcttggg gatcttgcgc cagcgtctct tgtcccagcg gcaaagcagc
     2161 agcaggcgaa aggtgtcccg gaaggctttg ttgcagagtg cgtagcacat ggggttgatg
     2221 gtgctgttga cgtagcacag ccagtagccc agctcccaca gggtctcggg aacacagtcc
     2281 ttgcagaagg tggacaccag caccatgatg ttgtacggtg tccaggtgag gatgaaggcc
     2341 aggaggatgg cactcagggt ccgagccgcc ttcttctcct tgaccagcga gaaggtcttc
     2401 cgcttggcca gctgctcctt tccacggggc ttctggccct tgccagctcg atcacgccct
     2461 ttcttagtcg gcctcttgac tgtatttggg gagctccgtg ggggctgctt ggtgggggcc
     2521 tgtgcctcgg ggtccaccat tggcatcttg atcaccactt cggagccagg ctcctctccc
     2581 tctgaggatg tgagggactc catggagcct tcgtcctctt cctcttcttc cttccagctg
     2641 taggcctgca gcagcctggg ggcccggcag cagcgacagc agcggcctgg aggagtctct
     2701 ggtgagccct cagcccctgg ctgagacctc tctgagctgc tgctgctgcc accccctttg
     2761 cctggcgtct cggagccctg aagggctgcc agctcccgtg ctcggttctc tgtctcccgg
     2821 tagatgcgcc agtagagcgt gcacatgact gtgacaggga ggtagaaggc agccatggct
     2881 gtgccaaagg tgatgatggg ctgggagagg aactggatgt agcactgccc agctagcact
     2941 gtccgctccc ctaccaggta ctgccagaag aggatggctg gggcccagag cacaaaggaa
     3001 accagccagg ccaggccgat catcagagct gcccggcggg gtgtgcgctt ggcacggtag
     3061 ctcaggggcc gagtcacgga gaagtagcgg tcaaagctga tgagcagcag attcatgacg
     3121 gaggcattgc tggccacata gtccagggcc agccagaggt cacaagccag cgtgcccaga
     3181 gcccagtggc ccatgagcag gtacgtggta tagaggttca tggagaaggt accgatgatg
     3241 aggtcagcac aggccaggct cagcaggaag tagttattga ctgtcttgag ctccgtgttg
     3301 accttgaaag agatgagtac cagcaggttg cctgtcactg tggctagcga caggaggccc
     3361 gtggtgatcc caatgaaggc cacttgccag ggaccctttc ctggtgccag gacggtgatg
     3421 ttggggctga cagcaggtgg ggctgaagtg ttcatggcga tcgctagccc tatagtgagt
     3481 cgtattaata ctctagcctt aagagctgta attgaactgg gagtggacac ctgtggagag
     3541 aaaggcaaag tggatgtcag taagaccaat aggtgcctat cagaaacgca agagtcttct
     3601 ctgtctcgac aagcccagtt tctattggtc tccttaaacc tgtcttgtaa ccttgatact
     3661 tacctgccca gtgcctcacg accaacttct gcagcttaag ttcgagactg ttgtgtcaga
     3721 agcactgact gcgttagcaa tttaactgtg ataaactacc gcaataaagc ttctagtgat
     3781 ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     3841 catttgcgtc aacggggcgg ggttattacg acattttgga aagtcccgtt gattttggtg
     3901 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     3961 ccgctatcca cgcccattgg tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     4021 taatacgtag atgtactgcc aagtaggaaa gtcccgtaag gtcatgtact gggcataatg
     4081 ccaggcgggc catttaccgt cattgacgtc aatagggggc ggacttggca tatgatacac
     4141 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     4201 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     4261 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     4321 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtgg cgcgccataa
     4381 tcctaggctc tcgtacgaag tcagctgata acgtcgacgt attgcggccg cggatcataa
     4441 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4501 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4561 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4621 cgacgtctta attaacgcgt cttgcgcagc accatggcct gaaataacct ctgaaagagg
     4681 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4741 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4801 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4861 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4921 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4981 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5041 cctaggcttt tgcaaaaagc tcgattcttc tgacactagc gccaccatgg ccaagccttt
     5101 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     5161 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     5221 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     5281 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     5341 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     5401 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     5461 ctctggttat gtgtgggagg gctaaactct agccatcacg atggccgcaa taaaatatct
     5521 ttattttcat tacatctgtg tgttggtttt ttgtgtgacg attccggact ataaaaggat
     5581 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5641 ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
