LOCUS       sequence-479831-        5303 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express DAP12-LgBiT 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 5303)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5303)
  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            "Unpublished"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: DAP12-LgBiT BiBiT Ready Vector
FEATURES             Location/Qualifiers
     source          1..5303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1183..1201)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1184..1201)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1321..1624
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1779..1799
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1832..1879
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1911..2705
                     /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(1965..1984)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2575..2594
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      2922..3467
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3368..3387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3584..3601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3751..3799
                     /note="synthetic polyadenylation signal"
     CDS             complement(3823..4860)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3836..3844
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(4891..5248)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4905..5040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5057..5248
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5205..5225
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttagct gttgatggtt actcggaaca
      301 gcatggagcc gtcgggggtg atcaggcgct cgtcgataat tttgttgccg ttccacaggg
      361 tccctgttac agtgatcttt ttgccgtcga acacggcgat gccttcatac ggccgtccga
      421 aatagttcag catgttcggc gtaaccccgt cgattaccag tgtgccatag ggcaggatca
      481 ccttaaagtg atgatcatcc acagggtaca ccaccttaaa cacctcttcg atctgggcca
      541 tttggtcggc gctcagacct tcatacggga tgatgacatg gatgtcgatc ttcagggcat
      601 tttcaccgct ccggacaatc ctttggatcg gagttacgga cacggcgaga ttctgcagca
      661 aactggacac acctccctgt tcaaggactt ggtccaggtt gtaggcggct gtctgttccc
      721 agtccccaac gaaatcttcg agtgtgaaga caccgctcga gcctccacct ccgctcccgc
      781 caccacctga actgccttga gaaactttgt aatacggcct ctgtgtgttg aggtcgctgt
      841 agacatccga cctctgaccc tggagctcct gataaggcga ctcggtctca gtgatacgct
      901 gtttccgggt cgctgcctcc gcagcccctc gcccccgagg gaccagccgg cccaggaagt
      961 acacggccag ggcaatgagc actgtcagca ccaggtctcc catcacgatc cctgccagca
     1021 cgcccgggct caccgtagag caactgcaat cgctctgggc ctgggcctgg acaggacgga
     1081 gaccacttac agccagcagg agaggcagga gcaggagcct gctgcagggt tcaagtcccc
     1141 ccatggcgat cgctagctgc aaaaagaaca agtagcttgt attctatagt gtcacctaaa
     1201 tgggcgatct gacggttcac taaaccagct ctgcttatat aggcctccct ggcgcgccac
     1261 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     1321 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     1381 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     1441 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     1501 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     1561 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1621 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
     1681 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1741 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
     1801 acggtgggag gtcaaatcga attctaatac tgttgacaat taatcatcgg catagtatat
     1861 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     1921 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     1981 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2041 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2101 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     2161 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     2221 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     2281 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     2341 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     2401 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     2461 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     2521 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     2581 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     2641 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     2701 tctgattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     2761 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtcaa acatgagaat
     2821 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
     2881 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
     2941 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
     3001 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
     3061 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     3121 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     3181 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     3241 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     3301 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     3361 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     3421 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     3481 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     3541 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     3601 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     3661 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     3721 cactccgcta gcgctgatgt ccggaatcgt cacacaaaaa accaacacac agatgtaatg
     3781 aaaataaaga tattttattg cggccatcgt gatggctaga gtctaaactc gacctacctc
     3841 cttagcgcgc ggtcgtgtac tgggccggcg gttgccgctg tcggccagca cctcgacgca
     3901 gccgtcggtc catacggctg cgctccggcg agcgatttgt gtgcgaccga cggtgccggc
     3961 cccgctgcgg acgatggcat cgcagcggcc ttgagcccag gcagcatcgt cgaagttgcc
     4021 gtccacgagg ctctggtaga gctgatccag gccgatgcgc agcatgtagg ctcgcagacg
     4081 agggctgccg gccagctcgg gatgccggcg ctcgaagtag cgagtctgct gctccatgca
     4141 agccagccag ggccgccaga agaagatgtt ggccacctcg tactgactgt ccccgaacat
     4201 agcttcggac cagtcgatta cggcggtgat gcggccgttg tcggtcagga cgttgttgct
     4261 gccgaagtcg gcatggacga ggtggcgcac ctcgggacag tcttcggccc acagcatcag
     4321 ttcgtccagg gcttgagcta cgctggcgga cacggtgtcg tccatcacgg tctgccagtg
     4381 gtagacatgg ggatcagcaa tggcgcaaat gaaatcccgc caggttgtgt actggccgat
     4441 gccttggggc ccgaatgggc cgaagccgct ggtttgactg aggtcggcgg cggcaatagc
     4501 atccatagct tcggcgacag gctgtaacac agcaggcagc tctgtttcgg ggaggtcttg
     4561 gagagtaacg ccttgggcgc gtctactgat gcagtatgtc aggctctcgc tgaactcgcc
     4621 gatgtccaac acttcgggga tgggtagtgc agcgctggcg aagtggcggt acacgtagcg
     4681 gtctttgtag aagccatcag cgcagctatt cacccgcagt acatagccgc gtccgccgac
     4741 atcgaagctg aaggctcggc tctcttcgcc ctccgacaac tgcatcaggt cgctcacact
     4801 gtcgaacttc tcgatgagaa atttttcaac gctggtagcg gtgagttcgg gcttcttcat
     4861 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     4921 tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa
     4981 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5041 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     5101 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     5161 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     5221 attccacagc tggttctttc cgcctcagaa ggtacctaac caagttcctc tttcagaggt
     5281 tatttcaggc catggtgctg cgc
//
