LOCUS       sequence-491033-        5714 bp    DNA     circular SYN 29-OCT-2025
DEFINITION  Express HiBiT-GLP1R 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 5714)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5714)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-GLP1R Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5714
                     /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(2657..2675)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2712..2833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2723..2742)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2777..2796
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3078..3435
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3078..3269
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3101..3121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3286..3421
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3450..3497
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3529..4323
                     /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(3583..3602)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4193..4212
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4387..4435
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4674..5262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4754..4773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5378..5661
                     /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 gcccgccccc agggtgccac tgtgtccctc tgggagacgg
     1261 tgcagaaatg gcgagaatac cgacgccagt gccagcgctc cctgactgag gatccacctc
     1321 ctgccacaga cttgttctgc aaccggacct tcgatgaata cgcctgctgg ccagatgggg
     1381 agccaggctc gttcgtgaat gtcagctgcc cctggtacct gccctgggcc agcagtgtgc
     1441 cgcagggcca cgtgtaccgg ttctgcacag ctgaaggcct ctggctgcag aaggacaact
     1501 ccagcctgcc ctggagggac ttgtcggagt gcgaggagtc caagcgaggg gaaagaagct
     1561 ccccggagga gcagctcctg ttcctctaca tcatctacac ggtgggctac gcactctcct
     1621 tctctgctct ggttatcgcc tctgcgatcc tcctcggctt cagacacctg cactgcacca
     1681 ggaactacat ccacctgaac ctgtttgcat ccttcatcct gcgagcattg tccgtcttca
     1741 tcaaggacgc agccctgaag tggatgtata gcacagccgc ccagcagcac cagtgggatg
     1801 ggctcctctc ctaccaggac tctctgagct gccgcctggt gtttctgctc atgcagtact
     1861 gtgtggcggc caattactac tggctcttgg tggagggcgt gtacctgtac acactgctgg
     1921 ccttctcggt cttatctgag caatggatct tcaggctcta cgtgagcata ggctggggtg
     1981 ttcccctgct gtttgttgtc ccctggggca ttgtcaagta cctctatgag gacgagggct
     2041 gctggaccag gaactccaac atgaactact ggctcattat ccggctgccc attctctttg
     2101 ccattggggt gaacttcctc atctttgttc gggtcatctg catcgtggta tccaaactga
     2161 aggccaatct catgtgcaag acagacatca aatgcagact tgccaagtcc acgctgacac
     2221 tcatccccct gctggggact catgaggtca tctttgcctt tgtgatggac gagcacgccc
     2281 gggggaccct gcgcttcatc aagctgttta cagagctctc cttcacctcc ttccaggggc
     2341 tgatggtggc catattatac tgctttgtca acaatgaggt ccagctggaa tttcggaaga
     2401 gctgggagcg ctggcggctt gagcacttgc acatccagag ggacagcagc atgaagcccc
     2461 tcaagtgtcc caccagcagc ctgagcagtg gagccacggc gggcagcagc atgtacacag
     2521 ccacttgcca ggcctcctgc agcgtttaaa cgaattcggg ctcggtaccc ggggatcctc
     2581 tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     2641 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     2701 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2761 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2821 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2881 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     2941 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3001 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     3061 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3121 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3181 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3241 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3301 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3361 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3421 ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca tagtatagcg
     3481 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     3541 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3601 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3661 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3721 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3781 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3841 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3901 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3961 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4021 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4081 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4141 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4201 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4261 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4321 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     4381 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4441 atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     4501 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt atcagctcac
     4561 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4621 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4921 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4981 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5041 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5101 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5161 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5221 gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt
     5281 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5341 ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct
     5401 ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg
     5461 ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag
     5521 gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga
     5581 tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact
     5641 gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta
     5701 gaggatcgag atct
//
