LOCUS       sequence-480846-        8492 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  Express LgBiT-JAK2 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 8492)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8492)
  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            "Unpublished"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: LgBiT-JAK2 Vector
FEATURES             Location/Qualifiers
     source          1..8492
                     /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"
     primer_bind     275..293
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(4372..4390)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4373..4390)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        4510..4813
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4968..4988
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        5021..5068
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5100..5894
                     /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(5154..5173)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5764..5783
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      6111..6656
                     /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     6557..6576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6773..6790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6940..6988
                     /note="synthetic polyadenylation signal"
     CDS             complement(7012..8049)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             7025..7033
                     /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(8080..8437)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8094..8229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        8246..8437
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     8394..8414
                     /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 ccccaagggg ttatgctagt tattgctcag cggtggcagc
      301 agccaactca gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg
      361 tcgactctag aggatccccg ggtaccgagc ccgaattcgt ttaaactcca gccatgttat
      421 cccttatttg atccactcga agagctagat ccctaaagga ggggcgttga tttacattat
      481 tgttccagca ttctgtcatg atcatataga tctcatctgg gcatccatct ggtcttggta
      541 atcttccatt attcttcaaa agttctatca aatggaacac gatcatctgt ccttgtttgt
      601 cattgccaat catacgcata aattccgctg gtggactttt actcttctca atgtatgtga
      661 aaagttcata cagaaccact ccaaagctcc aaacatctga ggccacagaa aacttgctct
      721 ctgtcagtga ttctggagca taccagaata tgggactttc accaggttct tttactttat
      781 agtattcttt gtcttgtggc aagactttgg ttaacccaaa atctccaatt ttaactctgt
      841 tctcgttctc caccaatata tttctcgttg ccagatccct gtggatatac ctttttgtac
      901 caagatactc catacccttg catatctgag atgtgtactg cagaagtttt atgtgatcta
      961 tccgttcttt atgtttttga agatagtctc gtaaacttcc atatggtaaa tattccataa
     1021 ttaattttag attacgccga ccagcactgt agcacactcc cttgtacttt acaatgttgt
     1081 catgctgtag ggatttcagg atttcaattt ccctttcaaa gtctcttagg tgctcttcag
     1141 tactatgctg aagctttttt acagcgacca cctccccagt gttgtcctgt agagggtcat
     1201 accggcacat ctccacactc ccaaaattac ccttgccaag ttgctgtaga aatttcaaat
     1261 gtctctcttc aaactgtgta ggatcccggt cttcaaaggc accagaaaac cctagggcac
     1321 ctatcctcat atttggtaac atgtcatttt ctgttaatag ttcataatct ggagtaaaca
     1381 aactgttaag atctcgtatg atggctctga aagaaggcct gaaatctggt tcataatcca
     1441 tacaattatt tataaggttt gctaattctg cccactttgg tgcaggaagc tgatgcctat
     1501 cttcataaaa ttgtagcttt ctttgagaat ccagagcact tagaggttta tctcctccac
     1561 tgcagatttc ccacaaagtg gtaccaaaac tccatttgtc tgttgccaaa tttaaatttt
     1621 taggattttc aatgcattca ggtggtaccc atggtattct ctcctgaaga atgtcctttg
     1681 gcaaaactgt aatactaatg ccaggatcac taagtttgat gaaaggagga tttcctgtct
     1741 tcctgtcttc ttctctgata agcagaatat ttttggcaca tacattccca tgaataaggg
     1801 tgttttcttc tagaaaatgc atggcccatg ccaactgttt agcaacttca agtttccata
     1861 atatatttat acaattttta ttctttttca gatatgtatc tagtgatcca aattttacaa
     1921 actcctgaac cagaatattc tcgtctccac agacacatac tccataattt aaaaccaaat
     1981 gcttgtgaga aagcttgctc atcatacttg ctgcttcaaa gaaagactct gaatagtttc
     2041 tgtgtgcttt atccagaact tttaaaagaa cttctgtttc atgcagttga ccgtagtctc
     2101 ctacttctct tcgtacgcct ttaaaaatct ttgtaaaagt gccttggcca aggctttcat
     2161 taaatatcaa atcttcattt ctgattttgt gaaacaccat ttggttcata tgagtaggcc
     2221 tctgtaatgt tggtgaggtt ggtacatcag aaacaccatt cgttctgaag actagaaggt
     2281 ttgatttatc ttttggcttt gggggacagc atttagtaaa ctggaaaatt atattgtctg
     2341 agcgaacagt ttccatctgg taacaattca aaagatcttt aagactgctg aagttcttct
     2401 ttgtcccact gaggttgtac tcttcattct cattttttgt aatcaaacag tgtttatatt
     2461 caatgacatt ttctcgctcg acagcaaaag tcaaaaaata tttattaaag tccttagggc
     2521 tgcatcgaag tacatacagt ccagtctgat tacctgcttt cttcagttta ctaatggcaa
     2581 aatccatcga aattgggcca tgacagttgc tttgtatatt ttcaagcacg gctggaggtg
     2641 ctacttcttt acagaggtaa tgatgtgcat ctgcagttaa tctataatat ccatcaatta
     2701 atgacacgaa agacaaagct tcccttaatg agctaagttc aatttccaga tttttaccat
     2761 cttgcttatg gatagttaca actcggcttt catttgaacc ctcttggttt gcttgcttaa
     2821 tactgacatc aataatatta ggaaaatcgc aatataactg taaatcctgt tctgtcagtg
     2881 tctcactttc tttatgtttc cctcttgacc actgaattcc accgtttcca gttattataa
     2941 tggttgcaaa aatctcctca cctgaaggac cacttccagg ttcttttact tcaaatttct
     3001 ctgtgtagaa ggcagactgc agagtttcca gatttataag atacttaagt ttcaagtttc
     3061 tggcagtggc tttgcattgg ctgaattgct gaataaatct gcgaaatctg taccttattc
     3121 gcttccttgt caaaatatga tagtcttgga tctttgctcg aatacatttt ggtaagaatg
     3181 tcttgtagct gatagagtta tagatggcca gtggggtttg atcgttttct ttggctattc
     3241 tcatcatatc taacactgcc atcccaagac attcttcctg tgtttcatga gtcacaggta
     3301 cttttatcca tccatgcaca aaatcatgcc gccactgagc aaagaggtaa gacatgacaa
     3361 agtcatcaag aagaggagct tcagcacctc gagatattcc atgccgatag gctctgttgc
     3421 tgccactgca ataccaacga ggaaagtaaa atcttattct gtagagtaca ttatgcctgg
     3481 ttgactcatc tatatggaag acatggttgg gtggatacca gatcctttct gtttcactca
     3541 ttaaagcaaa catattatga tacacaggtg tgataccaca agctttagaa gcagcaatac
     3601 agatttcttc tgcaacatac tccccagatg gaaaggtcag ataatctgcc tcagatttcc
     3661 caagggaatg gtaaagatac acctgaagaa ctggatctat ttgcttcata gaattggcat
     3721 ttccagaaat atcaccattc tgatatatag aagaggtgga tgttccctcc atttctgtca
     3781 tcgtaaggca ggccattccc atggcgatcg caccgctcga gcctccacct ccgctcccgc
     3841 caccaccgga gctcccactg ttgatggtta ctcggaacag catggagccg tcgggggtga
     3901 tcaggcgctc gtcgataatt ttgttgccgt tccacagggt ccctgttaca gtgatctttt
     3961 tgccgtcgaa cacggcgatg ccttcatacg gccgtccgaa atagttcagc atgttcggcg
     4021 taaccccgtc gattaccagt gtgccatagg gcaggatcac cttaaagtga tgatcatcca
     4081 cagggtacac caccttaaac acctcttcga tctgggccat ttggtcggcg ctcagacctt
     4141 catacgggat gatgacatgg atgtcgatct tcagggcatt ttcaccgctc cggacaatcc
     4201 tttggatcgg agttacggac acggcgagat tctgcagcaa actggacaca cctccctgtt
     4261 caaggacttg gtccaggttg taggcggctg tctgttccca gtccccaacg aaatcttcga
     4321 gtgtgaagac catggtgagc gctagctgca aaaagaacaa gtagcttgta ttctatagtg
     4381 tcacctaaat gggcgatctg acggttcact aaaccagctc tgcttatata ggcctccctg
     4441 gcgcgccact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     4501 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     4561 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     4621 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     4681 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     4741 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     4801 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     4861 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     4921 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     4981 taggcgtgta cggtgggagg tcaaatcgaa ttctaatact gttgacaatt aatcatcggc
     5041 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     5101 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5161 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5221 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5281 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5341 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5401 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5461 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5521 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5581 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5641 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5701 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5761 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5821 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5881 acgagttctt ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc
     5941 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtcaaa
     6001 catgagaatt acaacttata tcgtatgggg ctgacttcag gtgctacatt tgaagagata
     6061 aattgcactg aaatctagaa atattttatc tgattaataa gatgatcttc ttgagatcgt
     6121 tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt
     6181 tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg aggagcgcag
     6241 tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa gactaactcc
     6301 tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt ccgggttgga
     6361 ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg gttcgtgcat
     6421 acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca
     6481 aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca
     6541 cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6601 actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg
     6661 gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt ccaggaaatc
     6721 tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc
     6781 agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca ccggtgcagc
     6841 cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc aacatagtaa
     6901 gccagtatac actccgctag cgctgatgtc cggaatcgtc acacaaaaaa ccaacacaca
     6961 gatgtaatga aaataaagat attttattgc ggccatcgtg atggctagag tctaaactcg
     7021 acctacctcc ttagcgcgcg gtcgtgtact gggccggcgg ttgccgctgt cggccagcac
     7081 ctcgacgcag ccgtcggtcc atacggctgc gctccggcga gcgatttgtg tgcgaccgac
     7141 ggtgccggcc ccgctgcgga cgatggcatc gcagcggcct tgagcccagg cagcatcgtc
     7201 gaagttgccg tccacgaggc tctggtagag ctgatccagg ccgatgcgca gcatgtaggc
     7261 tcgcagacga gggctgccgg ccagctcggg atgccggcgc tcgaagtagc gagtctgctg
     7321 ctccatgcaa gccagccagg gccgccagaa gaagatgttg gccacctcgt actgactgtc
     7381 cccgaacata gcttcggacc agtcgattac ggcggtgatg cggccgttgt cggtcaggac
     7441 gttgttgctg ccgaagtcgg catggacgag gtggcgcacc tcgggacagt cttcggccca
     7501 cagcatcagt tcgtccaggg cttgagctac gctggcggac acggtgtcgt ccatcacggt
     7561 ctgccagtgg tagacatggg gatcagcaat ggcgcaaatg aaatcccgcc aggttgtgta
     7621 ctggccgatg ccttggggcc cgaatgggcc gaagccgctg gtttgactga ggtcggcggc
     7681 ggcaatagca tccatagctt cggcgacagg ctgtaacaca gcaggcagct ctgtttcggg
     7741 gaggtcttgg agagtaacgc cttgggcgcg tctactgatg cagtatgtca ggctctcgct
     7801 gaactcgccg atgtccaaca cttcggggat gggtagtgca gcgctggcga agtggcggta
     7861 cacgtagcgg tctttgtaga agccatcagc gcagctattc acccgcagta catagccgcg
     7921 tccgccgaca tcgaagctga aggctcggct ctcttcgccc tccgacaact gcatcaggtc
     7981 gctcacactg tcgaacttct cgatgagaaa tttttcaacg ctggtagcgg tgagttcggg
     8041 cttcttcatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     8101 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     8161 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     8221 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     8281 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     8341 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     8401 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     8461 ttcagaggtt atttcaggcc atggtgctgc gc
//
