LOCUS       sequence-480845-        8360 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  Express JAK2-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 8360)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8360)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: JAK2-LgBiT Vector
FEATURES             Location/Qualifiers
     source          1..8360
                     /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(4240..4258)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4241..4258)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        4378..4681
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4836..4856
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        4889..4936
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4968..5762
                     /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(5022..5041)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5632..5651
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5979..6524
                     /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     6425..6444
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6641..6658
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6808..6856
                     /note="synthetic polyadenylation signal"
     CDS             complement(6880..7917)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             6893..6901
                     /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(7948..8305)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7962..8097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8114..8305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8262..8282
                     /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 gaaactccag ccatgttatc ccttatttga tccactcgaa
      841 gagctagatc cctaaaggag gggcgttgat ttacattatt gttccagcat tctgtcatga
      901 tcatatagat ctcatctggg catccatctg gtcttggtaa tcttccatta ttcttcaaaa
      961 gttctatcaa atggaacacg atcatctgtc cttgtttgtc attgccaatc atacgcataa
     1021 attccgctgg tggactttta ctcttctcaa tgtatgtgaa aagttcatac agaaccactc
     1081 caaagctcca aacatctgag gccacagaaa acttgctctc tgtcagtgat tctggagcat
     1141 accagaatat gggactttca ccaggttctt ttactttata gtattctttg tcttgtggca
     1201 agactttggt taacccaaaa tctccaattt taactctgtt ctcgttctcc accaatatat
     1261 ttctcgttgc cagatccctg tggatatacc tttttgtacc aagatactcc atacccttgc
     1321 atatctgaga tgtgtactgc agaagtttta tgtgatctat ccgttcttta tgtttttgaa
     1381 gatagtctcg taaacttcca tatggtaaat attccataat taattttaga ttacgccgac
     1441 cagcactgta gcacactccc ttgtacttta caatgttgtc atgctgtagg gatttcagga
     1501 tttcaatttc cctttcaaag tctcttaggt gctcttcagt actatgctga agctttttta
     1561 cagcgaccac ctccccagtg ttgtcctgta gagggtcata ccggcacatc tccacactcc
     1621 caaaattacc cttgccaagt tgctgtagaa atttcaaatg tctctcttca aactgtgtag
     1681 gatcccggtc ttcaaaggca ccagaaaacc ctagggcacc tatcctcata tttggtaaca
     1741 tgtcattttc tgttaatagt tcataatctg gagtaaacaa actgttaaga tctcgtatga
     1801 tggctctgaa agaaggcctg aaatctggtt cataatccat acaattattt ataaggtttg
     1861 ctaattctgc ccactttggt gcaggaagct gatgcctatc ttcataaaat tgtagctttc
     1921 tttgagaatc cagagcactt agaggtttat ctcctccact gcagatttcc cacaaagtgg
     1981 taccaaaact ccatttgtct gttgccaaat ttaaattttt aggattttca atgcattcag
     2041 gtggtaccca tggtattctc tcctgaagaa tgtcctttgg caaaactgta atactaatgc
     2101 caggatcact aagtttgatg aaaggaggat ttcctgtctt cctgtcttct tctctgataa
     2161 gcagaatatt tttggcacat acattcccat gaataagggt gttttcttct agaaaatgca
     2221 tggcccatgc caactgttta gcaacttcaa gtttccataa tatatttata caatttttat
     2281 tctttttcag atatgtatct agtgatccaa attttacaaa ctcctgaacc agaatattct
     2341 cgtctccaca gacacatact ccataattta aaaccaaatg cttgtgagaa agcttgctca
     2401 tcatacttgc tgcttcaaag aaagactctg aatagtttct gtgtgcttta tccagaactt
     2461 ttaaaagaac ttctgtttca tgcagttgac cgtagtctcc tacttctctt cgtacgcctt
     2521 taaaaatctt tgtaaaagtg ccttggccaa ggctttcatt aaatatcaaa tcttcatttc
     2581 tgattttgtg aaacaccatt tggttcatat gagtaggcct ctgtaatgtt ggtgaggttg
     2641 gtacatcaga aacaccattc gttctgaaga ctagaaggtt tgatttatct tttggctttg
     2701 ggggacagca tttagtaaac tggaaaatta tattgtctga gcgaacagtt tccatctggt
     2761 aacaattcaa aagatcttta agactgctga agttcttctt tgtcccactg aggttgtact
     2821 cttcattctc attttttgta atcaaacagt gtttatattc aatgacattt tctcgctcga
     2881 cagcaaaagt caaaaaatat ttattaaagt ccttagggct gcatcgaagt acatacagtc
     2941 cagtctgatt acctgctttc ttcagtttac taatggcaaa atccatcgaa attgggccat
     3001 gacagttgct ttgtatattt tcaagcacgg ctggaggtgc tacttcttta cagaggtaat
     3061 gatgtgcatc tgcagttaat ctataatatc catcaattaa tgacacgaaa gacaaagctt
     3121 cccttaatga gctaagttca atttccagat ttttaccatc ttgcttatgg atagttacaa
     3181 ctcggctttc atttgaaccc tcttggtttg cttgcttaat actgacatca ataatattag
     3241 gaaaatcgca atataactgt aaatcctgtt ctgtcagtgt ctcactttct ttatgtttcc
     3301 ctcttgacca ctgaattcca ccgtttccag ttattataat ggttgcaaaa atctcctcac
     3361 ctgaaggacc acttccaggt tcttttactt caaatttctc tgtgtagaag gcagactgca
     3421 gagtttccag atttataaga tacttaagtt tcaagtttct ggcagtggct ttgcattggc
     3481 tgaattgctg aataaatctg cgaaatctgt accttattcg cttccttgtc aaaatatgat
     3541 agtcttggat ctttgctcga atacattttg gtaagaatgt cttgtagctg atagagttat
     3601 agatggccag tggggtttga tcgttttctt tggctattct catcatatct aacactgcca
     3661 tcccaagaca ttcttcctgt gtttcatgag tcacaggtac ttttatccat ccatgcacaa
     3721 aatcatgccg ccactgagca aagaggtaag acatgacaaa gtcatcaaga agaggagctt
     3781 cagcacctcg agatattcca tgccgatagg ctctgttgct gccactgcaa taccaacgag
     3841 gaaagtaaaa tcttattctg tagagtacat tatgcctggt tgactcatct atatggaaga
     3901 catggttggg tggataccag atcctttctg tttcactcat taaagcaaac atattatgat
     3961 acacaggtgt gataccacaa gctttagaag cagcaataca gatttcttct gcaacatact
     4021 ccccagatgg aaaggtcaga taatctgcct cagatttccc aagggaatgg taaagataca
     4081 cctgaagaac tggatctatt tgcttcatag aattggcatt tccagaaata tcaccattct
     4141 gatatataga agaggtggat gttccctcca tttctgtcat cgtaaggcag gccattccca
     4201 tggcgatcgc tagctgcaaa aagaacaagt agcttgtatt ctatagtgtc acctaaatgg
     4261 gcgatctgac ggttcactaa accagctctg cttatatagg cctccctggc gcgccactag
     4321 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     4381 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4441 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4501 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4561 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4621 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4681 ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
     4741 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4801 ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg
     4861 gtgggaggtc aaatcgaatt ctaatactgt tgacaattaa tcatcggcat agtatatcgg
     4921 catagtataa tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag
     4981 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5041 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5101 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5161 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5221 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5281 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5341 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5401 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5461 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5521 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5581 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5641 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5701 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5761 gattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat
     5821 acggttatcc acagaatcag gggataacgc aggaaagaac atgtcaaaca tgagaattac
     5881 aacttatatc gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa
     5941 atctagaaat attttatctg attaataaga tgatcttctt gagatcgttt tggtctgcgc
     6001 gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc
     6061 tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc accaaaactt
     6121 gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc taaatcaatt
     6181 accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact caagacgata
     6241 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac agtccagctt
     6301 ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa cgcggccata
     6361 acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg agggagccgc
     6421 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
     6481 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
     6541 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
     6601 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
     6661 agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct
     6721 gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac
     6781 tccgctagcg ctgatgtccg gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa
     6841 ataaagatat tttattgcgg ccatcgtgat ggctagagtc taaactcgac ctacctcctt
     6901 agcgcgcggt cgtgtactgg gccggcggtt gccgctgtcg gccagcacct cgacgcagcc
     6961 gtcggtccat acggctgcgc tccggcgagc gatttgtgtg cgaccgacgg tgccggcccc
     7021 gctgcggacg atggcatcgc agcggccttg agcccaggca gcatcgtcga agttgccgtc
     7081 cacgaggctc tggtagagct gatccaggcc gatgcgcagc atgtaggctc gcagacgagg
     7141 gctgccggcc agctcgggat gccggcgctc gaagtagcga gtctgctgct ccatgcaagc
     7201 cagccagggc cgccagaaga agatgttggc cacctcgtac tgactgtccc cgaacatagc
     7261 ttcggaccag tcgattacgg cggtgatgcg gccgttgtcg gtcaggacgt tgttgctgcc
     7321 gaagtcggca tggacgaggt ggcgcacctc gggacagtct tcggcccaca gcatcagttc
     7381 gtccagggct tgagctacgc tggcggacac ggtgtcgtcc atcacggtct gccagtggta
     7441 gacatgggga tcagcaatgg cgcaaatgaa atcccgccag gttgtgtact ggccgatgcc
     7501 ttggggcccg aatgggccga agccgctggt ttgactgagg tcggcggcgg caatagcatc
     7561 catagcttcg gcgacaggct gtaacacagc aggcagctct gtttcgggga ggtcttggag
     7621 agtaacgcct tgggcgcgtc tactgatgca gtatgtcagg ctctcgctga actcgccgat
     7681 gtccaacact tcggggatgg gtagtgcagc gctggcgaag tggcggtaca cgtagcggtc
     7741 tttgtagaag ccatcagcgc agctattcac ccgcagtaca tagccgcgtc cgccgacatc
     7801 gaagctgaag gctcggctct cttcgccctc cgacaactgc atcaggtcgc tcacactgtc
     7861 gaacttctcg atgagaaatt tttcaacgct ggtagcggtg agttcgggct tcttcatggt
     7921 ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     7981 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     8041 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     8101 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     8161 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     8221 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     8281 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     8341 ttcaggccat ggtgctgcgc
//
