LOCUS       sequence-473381-        5628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Beclin-1-HiBiT in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5628)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5628)
  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            "Unpublished"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: Beclin-1-HiBiT Vector (CMV)
FEATURES             Location/Qualifiers
     source          1..5628
                     /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(2571..2589)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2626..2747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2637..2656)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2691..2710
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2992..3349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2992..3183
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3015..3035)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3200..3335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3364..3411
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3443..4237
                     /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(3497..3516)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4107..4126
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4301..4349
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4588..5176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4668..4687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5292..5575
                     /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 ggctagcgat cgccatggaa gggtctaaga
     1081 cgtccaacaa cagcaccatg caggtgagct tcgtgtgcca gcgctgcagc cagcccctga
     1141 aactggacac gagtttcaag atcctggacc gtgtcaccat ccaggaactc acagctccat
     1201 tacttaccac agcccaggcg aaaccaggag agacccagga ggaagagact aactcaggag
     1261 aggagccatt tattgaaact cctcgccagg atggtgtctc tcgcagattc atccccccag
     1321 ccaggatgat gtccacagaa agtgccaaca gcttcactct gattggggag gcatctgatg
     1381 gcggcaccat ggagaacctc agccgaagac tgaaggtcac tggggacctt tttgacatca
     1441 tgtcgggcca gacagatgtg gatcacccac tctgtgagga atgcacagat actcttttag
     1501 accagctgga cactcagctc aacgtcactg aaaatgagtg tcagaactac aaacgctgtt
     1561 tggagatctt agagcaaatg aatgaggatg acagtgaaca gttacagatg gagctaaagg
     1621 agctggcact agaggaggag aggctgatcc aggagctgga agacgtggaa aagaaccgca
     1681 agatagtggc agaaaatctc gagaaggtcc aggctgaggc tgagagactg gatcaggagg
     1741 aagctcagta tcagagagaa tacagtgaat ttaaacgaca gcagctggag ctggatgatg
     1801 agctgaagag tgttgaaaac cagatgcgtt atgcccagac gcagctggat aagctgaaga
     1861 aaaccaacgt ctttaatgca accttccaca tctggcacag tggacagttt ggcacaatca
     1921 ataacttcag gctgggtcgc ctgcccagtg ttcccgtgga atggaatgag attaatgctg
     1981 cttggggcca gactgtgttg ctgctccatg ctctggccaa taagatgggt ctgaaatttc
     2041 agagataccg acttgttcct tacggaaacc attcatatct ggagtctctg acagacaaat
     2101 ctaaggagct gccgttatac tgttctgggg ggttgcggtt tttctgggac aacaagtttg
     2161 accatgcaat ggtggctttc ctggactgtg tgcagcagtt caaagaagag gttgagaaag
     2221 gcgagacacg tttttgtctt ccctacagga tggatgtgga gaaaggcaag attgaagaca
     2281 caggaggcag tggcggctcc tattccatca aaacccagtt taactctgag gagcagtgga
     2341 caaaagctct caagttcatg ctgacgaatc ttaagtgggg tcttgcttgg gtgtcctcac
     2401 aattttataa caaagtttct caaggcagtt caggtggctc gagcggtgtg agcggctggc
     2461 ggctgttcaa gaagattagc taaggccgcg actctagagt cgacctgcag gcatgcaagc
     2521 tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2581 ataactagca taaccccttg gggcggccgc ttcgagcaga catgataaga tacattgatg
     2641 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2701 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2761 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta agcaagtaaa
     2821 acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac aatttcctga
     2881 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc
     2941 agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg
     3001 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3061 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3181 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3241 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3301 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaatta
     3361 actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     3421 aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc
     3481 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3541 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3601 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3661 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3721 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3781 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3841 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3901 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3961 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4021 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4081 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4141 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4201 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4261 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     4321 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct
     4381 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     4441 ctgcggactg gctttctacc cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4501 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4561 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4621 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4681 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4741 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4801 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4861 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4921 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4981 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5041 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5101 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5161 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5221 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5281 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     5341 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     5401 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     5461 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     5521 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     5581 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
