LOCUS       sequence-479916-        4955 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express SmBiT-BCL2L11 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 4955)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4955)
  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            "Unpublished"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: SmBiT-BCL2L11 Vector
FEATURES             Location/Qualifiers
     source          1..4955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2018..2139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2029..2048)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2254..2287)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2297..2344
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2376..3170
                     /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(2430..2449)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3040..3059
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3328..3916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3408..3427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4042..4090
                     /note="synthetic polyadenylation signal"
     CDS             complement(4114..4512)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4509..4518
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4543..4900)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4557..4692
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4709..4900
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4857..4877
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact atagggctag cgctcaccat ggtgaccggc
     1201 taccggctgt tcgaggagat tctcgggagc tccggtggtg gcgggagcgg aggtggaggc
     1261 tcgagcggtg cgatcgccat ggcaaagcaa ccttctgatg taagttctga gtgtgaccga
     1321 gaaggtagac aattgcagcc tgcggagagg cctccccagc tcagacctgg ggcccctacc
     1381 tccctacaga cagagccaca aggtaatcct gaaggcaatc acggaggtga aggggacagc
     1441 tgcccccacg gcagccctca gggcccgctg gccccacctg ccagccctgg cccttttgct
     1501 accagatccc cgcttttcat ctttatgaga agatcctccc tgctgtctcg atcctccagt
     1561 gggtatttct cttttgacac agacaggagc ccagcaccca tgagttgtga caaatcaaca
     1621 caaaccccaa gtcctccttg ccaggccttc aaccactatc tcagtgcaat ggcttccatg
     1681 aggcaggctg aacctgcaga tatgcgccca gagatatgga tcgcccaaga gttgcggcgt
     1741 attggagacg agtttaacgc ttactatgca aggagggtat ttttgaataa ttaccaagca
     1801 gccgaagacc acccacgaat ggttatctta cgactgttac gttacattgt ccgcctggtg
     1861 tggagaatgc atgtttaaac gaattcgggc tcggtacccg gggatcctct agagtcgacc
     1921 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1981 tgccaccgct gagcaaggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     2041 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2101 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     2161 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta aaacctctac
     2221 aaatgtggta aaatcttaat taagaattct aatataactt cgtatagcat acattatacg
     2281 aagttatgga tccactgttg acaattaatc atcggcatag tatatcggca tagtataata
     2341 cgacaaggtg aggaactaaa cccaggaggc agatcatgat tgaacaagat ggattgcacg
     2401 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2461 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2521 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     2581 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     2641 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     2701 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     2761 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     2821 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     2881 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     2941 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3001 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3061 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3121 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaaatga
     3181 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3901 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4021 ccttttatag tccggaatcg tcacacaaaa aaccaacaca cagatgtaat gaaaataaag
     4081 atattttatt gcggccatcg tgatggctag agtttagccc tcccacacat aaccagaggg
     4141 cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt
     4201 gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc
     4261 cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg cagcagcagc
     4321 agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata cattgacacc
     4381 agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt cttcagagat
     4441 ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt cttgagacaa
     4501 aggcttggcc atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc
     4561 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
     4621 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
     4681 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat
     4741 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
     4801 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc
     4861 taactgacac acattccaca gctggttctt tccgcctcag aaggtaccta accaagttcc
     4921 tctttcagag gttatttcag gccatggtgc tgcgc
//
