LOCUS       sequence-478461-        5774 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-CHRM1 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 5774)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5774)
  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            "Unpublished"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-CHRM1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5774
                     /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(2717..2735)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2772..2893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2837..2856
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3138..3495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3138..3329
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3161..3181)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3346..3481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3510..3557
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3589..4383
                     /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(3643..3662)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4253..4272
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4447..4495
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4734..5322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4814..4833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5438..5721
                     /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 gccatgaaca cttcagcccc acctgctgtc agccccaaca
     1261 tcaccgtcct ggcaccagga aagggtccct ggcaagtggc cttcattggg atcaccacgg
     1321 gcctcctgtc gctagccaca gtgacaggca acctgctggt actcatctct ttcaaggtca
     1381 acacggagct caagacagtc aataactact tcctgctgag cctggcctgt gctgacctca
     1441 tcatcggtac cttctccatg aacctctata ccacgtacct gctcatgggc cactgggctc
     1501 tgggcacgct ggcttgtgac ctctggctgg ccctggacta tgtggccagc aatgcctccg
     1561 tcatgaatct gctgctcatc agctttgacc gctacttctc cgtgactcgg cccctgagct
     1621 accgtgccaa gcgcacaccc cgccgggcag ctctgatgat cggcctggcc tggctggttt
     1681 cctttgtgct ctgggcccca gccatcctct tctggcagta cctggtaggg gagcggacag
     1741 tgctagctgg gcagtgctac atccagttcc tctcccagcc catcatcacc tttggcacag
     1801 ccatggctgc cttctacctc cctgtcacag tcatgtgcac gctctactgg cgcatctacc
     1861 gggagacaga gaaccgagca cgggagctgg cagcccttca gggctccgag acgccaggca
     1921 aagggggtgg cagcagcagc agctcagaga ggtctcagcc aggggctgag ggctcaccag
     1981 agactcctcc aggccgctgc tgtcgctgct gccgggcccc caggctgctg caggcctaca
     2041 gctggaagga agaagaggaa gaggacgaag gctccatgga gtccctcaca tcctcagagg
     2101 gagaggagcc tggctccgaa gtggtgatca agatgccaat ggtggacccc gaggcacagg
     2161 cccccaccaa gcagccccca cggagctccc caaatacagt caagaggccg actaagaaag
     2221 ggcgtgatcg agctggcaag ggccagaagc cccgtggaaa ggagcagctg gccaagcgga
     2281 agaccttctc gctggtcaag gagaagaagg cggctcggac cctgagtgcc atcctcctgg
     2341 ccttcatcct cacctggaca ccgtacaaca tcatggtgct ggtgtccacc ttctgcaagg
     2401 actgtgttcc cgagaccctg tgggagctgg gctactggct gtgctacgtc aacagcacca
     2461 tcaaccccat gtgctacgca ctctgcaaca aagccttccg ggacaccttt cgcctgctgc
     2521 tgctttgccg ctgggacaag agacgctggc gcaagatccc caagcgccct ggctccgtgc
     2581 accgcactcc ctcccgccaa tgcgtttaaa cgaattcggg ctcggtaccc ggggatcctc
     2641 tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     2701 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     2761 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2821 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2881 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2941 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     3001 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3061 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     3121 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3241 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3301 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3361 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3421 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3481 ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca tagtatatcg
     3541 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     3601 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3661 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3721 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3781 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3841 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3901 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3961 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4021 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4081 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4141 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4201 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4261 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4321 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4381 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     4441 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4501 atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     4561 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt atcagctcac
     4621 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4681 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4741 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4801 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4861 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4921 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4981 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5041 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5101 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5161 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5221 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5281 gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt
     5341 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5401 ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct
     5461 ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg
     5521 ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag
     5581 gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga
     5641 tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact
     5701 gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta
     5761 gaggatcgag atct
//
