LOCUS       sequence-473382-        5654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HiBiT-Beclin-1 in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5654)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5654)
  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            "Unpublished"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: HiBiT-Beclin-1 Vector (CMV)
FEATURES             Location/Qualifiers
     source          1..5654
                     /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(2597..2615)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2652..2773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2663..2682)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2717..2736
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3018..3375
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3018..3209
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3041..3061)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3226..3361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3390..3437
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3469..4263
                     /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(3523..3542)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4133..4152
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4327..4375
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4614..5202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4694..4713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5318..5601
                     /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 caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatggaag
     1141 ggtctaagac gtccaacaac agcaccatgc aggtgagctt cgtgtgccag cgctgcagcc
     1201 agcccctgaa actggacacg agtttcaaga tactggaccg tgtcaccatc caggaactca
     1261 cagctccatt acttaccaca gcccaggcga aaccaggaga gacccaggag gaagagacta
     1321 actcaggaga ggagccattt attgaaactc ctcgccagga tggtgtctct cgcagattca
     1381 tccccccagc caggatgatg tccacagaaa gtgccaacag cttcactctg attggggagg
     1441 catctgatgg cggcaccatg gagaacctca gccgaagact gaaggtcact ggggaccttt
     1501 ttgacatcat gtcgggccag acagatgtgg atcacccact ctgtgaggaa tgcacagata
     1561 ctcttttaga ccagctggac actcagctca acgtcactga aaatgagtgt cagaactaca
     1621 aacgctgttt ggagatctta gagcaaatga atgaggatga cagtgaacag ttacagatgg
     1681 agctaaagga gctggcacta gaggaggaga ggctgatcca ggagctggaa gacgtggaaa
     1741 agaaccgcaa gatagtggca gaaaatctcg agaaggtcca ggctgaggct gagagactgg
     1801 atcaggagga agctcagtat cagagagaat acagtgaatt taaacgacag cagctggagc
     1861 tggatgatga gctgaagagt gttgaaaacc agatgcgtta tgcccagacg cagctggata
     1921 agctgaagaa aaccaacgtc tttaatgcaa ccttccacat ctggcacagt ggacagtttg
     1981 gcacaatcaa taacttcagg ctgggtcgcc tgcccagtgt tcccgtggaa tggaatgaga
     2041 ttaatgctgc ttggggccag actgtgttgc tgctccatgc tctggccaat aagatgggtc
     2101 tgaaatttca gagataccga cttgttcctt acggaaacca ttcatatctg gagtctctga
     2161 cagacaaatc taaggagctg ccgttatact gttctggggg gttgcggttt ttctgggaca
     2221 acaagtttga ccatgcaatg gtggctttcc tggactgtgt gcagcagttc aaagaagagg
     2281 ttgagaaagg cgagacacgt ttttgtcttc cctacaggat ggatgtggag aaaggcaaga
     2341 ttgaagacac aggaggcagt ggcggctcct attccatcaa aacccagttt aactctgagg
     2401 agcagtggac aaaagctctc aagttcatgc tgacgaatct taagtggggt cttgcttggg
     2461 tgtcctcaca attttataac aaagtttaaa cgaattcggg ctcggtaccc ggggatcctc
     2521 tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     2581 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     2641 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2701 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2761 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2821 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     2881 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2941 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     3001 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3061 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3121 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3181 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3241 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3301 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3361 ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca tagtatatcg
     3421 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     3481 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3541 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3601 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3661 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3721 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3781 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3841 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3901 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3961 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4021 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4081 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4141 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4201 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4261 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     4321 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4381 atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     4441 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt atcagctcac
     4501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5041 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5161 gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt
     5221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5281 ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct
     5341 ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg
     5401 ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag
     5461 gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga
     5521 tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact
     5581 gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta
     5641 gaggatcgag atct
//
