LOCUS       sequence-479830-        6869 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Express Syk-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 6869)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6869)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Syk-LgBiT BiBiT Ready Vector
FEATURES             Location/Qualifiers
     source          1..6869
                     /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(2749..2767)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2750..2767)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2887..3190
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3345..3365
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3398..3445
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3477..4271
                     /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(3531..3550)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4141..4160
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      4488..5033
                     /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     4934..4953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5150..5167
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5317..5365
                     /note="synthetic polyadenylation signal"
     CDS             complement(5389..6426)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5402..5410
                     /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(6457..6814)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6471..6606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6623..6814
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6771..6791
                     /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 gaaacgttca ccacgtcata gtagtaattg cgcagccgca
      841 gttccactgc tgcgaatccg ggcctgtttt ccacatcgta tgtccagcac agattcatga
      901 gatcgtacat ctctcttgga caccctgcag ggcaccccat ccgctctcct ttctctaaca
      961 tagcggtgac ttcacttcct ttcatccctc gatatggctt ctgcccatag gagaatgctt
     1021 cccacatcaa cactccaaag ctccagacat cgcttttgct ggagaacttg tagtagttga
     1081 tgcattccgg agcgtaccac ttgacaggcc actttccatg ggtctgggcc ttgtagtagt
     1141 tttcatcagc acgcagtgct ttggaaagtc cgaaatcact gatcttggcg taatgttggg
     1201 taactagcaa cacatttctt gcagccagat ctctgtgcac aaaattgctc tcctccaagt
     1261 acttcatgcc catggaaacc tgatgaacca gttctatgat gttcttatcc ttgacatgtc
     1321 tgttctgctg caaatactta ttgaggggac caagttctgc catctccata accagcatcc
     1381 aggactcggc ctcgcatatc ccgatcatcc gcacgatgta cgggttgtcc agctgctgca
     1441 tgacatttgc ttctgctaat aactcatctt taagagcggg gtcattggcc tcgtttttca
     1501 gtattttcac agccacggtt ttcacaactt ttttcatttg gtagtagccc tttttcacag
     1561 ttccaaaatt accagagccc agttctttgt cttccagcgt cagcagcttt cggtccaggt
     1621 aaacctcctt gggcctgatc tcctcggggt ccgcgtaggg gctctcgtac acctctgtgt
     1681 ccatgggtag ggcttctctc tgggggcctt tgtctgcagc ccagggtgca agttctggct
     1741 catacggatt gaatgacaca gtactctctt gccggttccc ttgggcaggg gaggactttc
     1801 tgtggccagg ctttgggaag gagtatgatt tgattcttga gattattcca cccgctgacc
     1861 aagtcgcagg atgggaacct ggaagttgtg gacggcctcc aaaattaaca tttccctgtg
     1921 tgccgatttt ttgacatggg acagtaagaa ctcttaacaa accatctgct ttataagaat
     1981 aatgctcgac tagctgccag agcgtgtcga acttctttcc ctcggggatg gagagcttcc
     2041 ctgtcttgtc tttgtcgatg cgatagtgca gcaccttccc ttcgtgcagc aggcacaggg
     2101 cgtaggagcc gttgttgtct ctggctcgga tcaggaactt tccatttgtc tttgatccta
     2161 tcaggacaat ttgctcagat tcttcccgag agatttttcc atggaaccaa ggcatttttt
     2221 catgggctgt ggtagcgatc agcttctcca gctgaggctt ctgactgatg atggcctgct
     2281 ccagagcctg accctgcagg ttccatgtct gcttcacata ttccctgatg aggttttcct
     2341 tcaaatcctc aaagggccca gtcttgggct gcaccccttg gggccggttg aagggcttct
     2401 tgaggaggca gaccaggcca tcagactcct gggagtggta gtggcagagg tcggcggggc
     2461 tggcatgggt cctgccaccg gcgatggcgt aggtgccatt cagctcccgc tcgatggtgt
     2521 agtggtgtgc cttcctcccg tgggccacgg acagggcgaa gccacccagg tagttgcggc
     2581 tctggcgcag caaataaagc ccatcactca tgcccccctg gaccaggtaa tcttctgcct
     2641 cctcccgggt gatgttgccg aaaaagaagg gcaggtggtt ggcgctgtca gccatgccgc
     2701 tgctggccat ggcgatcgct agctgcaaaa agaacaagta gcttgtattc tatagtgtca
     2761 cctaaatggg cgatctgacg gttcactaaa ccagctctgc ttatataggc ctccctggcg
     2821 cgccactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2881 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2941 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3001 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3061 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3121 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3181 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
     3241 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     3301 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag
     3361 gcgtgtacgg tgggaggtca aatcgaattc taatactgtt gacaattaat catcggcata
     3421 gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga
     3481 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3541 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3601 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3661 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3721 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3781 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3841 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3901 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3961 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4021 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4081 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4141 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4201 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4261 agttcttctg attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa
     4321 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtcaaacat
     4381 gagaattaca acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat
     4441 tgcactgaaa tctagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt
     4501 ggtctgcgcg taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg
     4561 aaggttctct gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca
     4621 ccaaaacttg tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct
     4681 aaatcaatta ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc
     4741 aagacgatag ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca
     4801 gtccagcttg gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac
     4861 gcggccataa cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga
     4921 gggagccgcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
     4981 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
     5041 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc
     5101 gccccgttcg taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt
     5161 gagcgaggaa gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt
     5221 ttttctcctg ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc
     5281 agtatacact ccgctagcgc tgatgtccgg aatcgtcaca caaaaaacca acacacagat
     5341 gtaatgaaaa taaagatatt ttattgcggc catcgtgatg gctagagtct aaactcgacc
     5401 tacctcctta gcgcgcggtc gtgtactggg ccggcggttg ccgctgtcgg ccagcacctc
     5461 gacgcagccg tcggtccata cggctgcgct ccggcgagcg atttgtgtgc gaccgacggt
     5521 gccggccccg ctgcggacga tggcatcgca gcggccttga gcccaggcag catcgtcgaa
     5581 gttgccgtcc acgaggctct ggtagagctg atccaggccg atgcgcagca tgtaggctcg
     5641 cagacgaggg ctgccggcca gctcgggatg ccggcgctcg aagtagcgag tctgctgctc
     5701 catgcaagcc agccagggcc gccagaagaa gatgttggcc acctcgtact gactgtcccc
     5761 gaacatagct tcggaccagt cgattacggc ggtgatgcgg ccgttgtcgg tcaggacgtt
     5821 gttgctgccg aagtcggcat ggacgaggtg gcgcacctcg ggacagtctt cggcccacag
     5881 catcagttcg tccagggctt gagctacgct ggcggacacg gtgtcgtcca tcacggtctg
     5941 ccagtggtag acatggggat cagcaatggc gcaaatgaaa tcccgccagg ttgtgtactg
     6001 gccgatgcct tggggcccga atgggccgaa gccgctggtt tgactgaggt cggcggcggc
     6061 aatagcatcc atagcttcgg cgacaggctg taacacagca ggcagctctg tttcggggag
     6121 gtcttggaga gtaacgcctt gggcgcgtct actgatgcag tatgtcaggc tctcgctgaa
     6181 ctcgccgatg tccaacactt cggggatggg tagtgcagcg ctggcgaagt ggcggtacac
     6241 gtagcggtct ttgtagaagc catcagcgca gctattcacc cgcagtacat agccgcgtcc
     6301 gccgacatcg aagctgaagg ctcggctctc ttcgccctcc gacaactgca tcaggtcgct
     6361 cacactgtcg aacttctcga tgagaaattt ttcaacgctg gtagcggtga gttcgggctt
     6421 cttcatggtg gcgctagtgt cagaagaatc gagctttttg caaaagccta ggcctccaaa
     6481 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     6541 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     6601 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     6661 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     6721 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     6781 acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag ttcctctttc
     6841 agaggttatt tcaggccatg gtgctgcgc
//
