LOCUS       sequence-479800-        8412 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HaloTag-JAK3 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 8412)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8412)
  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            "Unpublished"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: HaloTag-JAK3 Vector
FEATURES             Location/Qualifiers
     source          1..8412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(5491..5509)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5546..5667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5557..5576)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5611..5630
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5912..6269
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5912..6103
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5935..5955)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6120..6255
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6284..6331
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6363..7157
                     /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(6417..6436)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7027..7046
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7221..7269
                     /note="synthetic polyadenylation signal"
     rep_origin      7426..7971
                     /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     7872..7891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8088..8105
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgccatggc acctccaagt gaagagacgc ccctgatccc
     2041 tcagcgttca tgcagcctct tgtccacgga ggctggtgcc ctgcatgtgc tgctgcccgc
     2101 tcggggcccc gggccccccc agcgcctatc tttctccttt ggggaccact tggctgagga
     2161 cctgtgcgtg caggctgcca aggccagcgg catcctgcct gtgtaccact ccctctttgc
     2221 tctggccacg gaggacctgt cctgctggtt ccccccgagc cacatcttct ccgtggagga
     2281 tgccagcacc caagtcctgc tgtacaggat tcgcttttac ttccccaatt ggtttgggct
     2341 ggagaagtgc caccgcttcg ggctacgcaa ggatttggcc agtgctatcc ttgacctgcc
     2401 agtcctggag cacctctttg cccagcaccg cagtgacctg gtgagtgggc gcctccccgt
     2461 gggcctcagt ctcaaggagc agggtgagtg tctcagcctg gccgtgttgg acctggcccg
     2521 gatggcgcga gagcaggccc agcggccggg agagctgctg aagactgtca gctacaaggc
     2581 ctgcctaccc ccaagcctgc gcgacctgat ccagggcctg agcttcgtga cgcggaggcg
     2641 tattcggagg acggtgcgca gagccctgcg ccgcgtggcc gcctgccagg cagaccggca
     2701 ctcgctcatg gccaagtaca tcatggacct ggagcggctg gatccagccg gggccgccga
     2761 gaccttccac gtgggcctcc ctggggccct tggtggccac gacgggctgg ggctgctccg
     2821 cgtggctggt gacggcggca tcgcctggac ccagggagaa caggaggtcc tccagccctt
     2881 ctgcgacttt ccagaaatcg tagacattag catcaagcag gccccgcgcg ttggcccggc
     2941 cggagagcac cgcctggtca ctgttaccag gacagacaac cagattttag aggccgagtt
     3001 cccagggctg cccgaggctc tgtcgttcgt ggcgctcgtg gacggctact tccggctgac
     3061 cacggactcc cagcacttct tctgcaagga ggtggcaccg ccgaggctgc tggaggaagt
     3121 ggccgagcag tgccacggcc ccatcactct ggactttgcc atcaacaagc tcaagactgg
     3181 gggctcacgt cctggctcct atgttctccg ccgcagcccc caggactttg acagcttcct
     3241 cctcactgtc tgtgtccaga acccccttgg tcctgattat aagggctgcc tcatccggcg
     3301 cagccccaca ggaaccttcc ttctggttgg cctcagccga ccccacagca gtcttcgaga
     3361 gctcctggca acctgctggg atggggggct gcacgtagat ggggtggcag tgaccctcac
     3421 ttcctgctgt atccccagac ccaaagaaaa gtccaacctg atcgtggtcc agagaggtca
     3481 cagcccaccc acatcatcct tggttcagcc ccaatcccaa taccagctga gtcagatgac
     3541 atttcacaag atccctgctg acagcctgga gtggcatgag aacctgggcc atgggtcctt
     3601 caccaagatt taccggggct gtcgccatga ggtggtggat ggggaggccc gaaagacaga
     3661 ggtgctgctg aaggtcatgg atgccaagca caagaactgc atggagtcat tcctggaagc
     3721 agcgagcttg atgagccaag tgtcgtaccg gcatctcgtg ctgctccacg gcgtgtgcat
     3781 ggctggagac agcaccatgg tgcaggaatt tgtacacctg ggggccatag acatgtatct
     3841 gcgaaaacgt ggccacctgg tgccagccag ctggaagctg caggtggtca aacagctggc
     3901 ctacgccctc aactatctgg aggacaaagg cctgccccat ggcaatgtct ctgcccggaa
     3961 ggtgctcctg gctcgggagg gggctgatgg gagcccgccc ttcatcaagc tgagtgaccc
     4021 tggggtcagc cccgctgtgt taagcctgga gatgctcacc gacaggatcc cctgggtggc
     4081 ccccgagtgt ctccgggagg cgcagacact tagcttggaa gctgacaagt ggggcttcgg
     4141 cgccacggtc tgggaagtgt ttagtggcgt caccatgccc atcagtgccc tggatcctgc
     4201 taagaaactc caattttatg aggaccggca gcagctgccg gcccccaagt ggacagagct
     4261 ggccctgctg attcaacagt gcatggccta tgagccggtc cagaggccct ccttccgagc
     4321 cgtcattcgt gacctcaata gcctcatctc ttcagactat gagctcctct cagaccccac
     4381 acctggtgcc ctggcacctc gtgatgggct gtggaatggt gcccagctct atgcctgcca
     4441 agaccccacg atcttcgagg agagacacct caagtacatc tcacagctgg gcaagggcaa
     4501 ctttggcagc gtggagctgt gccgctatga cccgctaggc gacaatacag gtgccctggt
     4561 ggccgtgaaa cagctgcagc acagcgggcc agaccagcag agggactttc agcgggagat
     4621 tcagatcctc aaagcactgc acagtgattt cattgtcaag tatcgtggtg tcagctatgg
     4681 cccgggccgc cagagcctgc ggctggtcat ggagtacctg cccagcggct gcttgcgcga
     4741 cttcctgcag cggcaccgcg cgcgcctcga tgccagccgc ctccttctct attcctcgca
     4801 gatctgcaag ggcatggagt acctgggctc ccgccgctgc gtgcaccgcg acctggccgc
     4861 ccgaaacatc ctcgtggaga gcgaggcaca cgtcaagatc gctgacttcg gcctagctaa
     4921 gctgctgccg cttgacaaag actactacgt ggtccgcgag ccaggccaga gccccatttt
     4981 ctggtatgcc cccgaatccc tctcggacaa catcttctct cgccagtcag acgtctggag
     5041 cttcggggtc gtcctgtacg agctcttcac ctactgcgac aaaagctgca gcccctcggc
     5101 cgagttcctg cggatgatgg gatgtgagcg ggatgtcccc gccctctgcc gcctcttgga
     5161 actgctggag gagggccaga ggctgccggc gcctcctgcc tgccctgctg aggttcacga
     5221 gctcatgaag ctgtgctggg cccctagccc acaggaccgg ccatcattca gcgccctggg
     5281 cccccagctg gacatgctgt ggagcggaag ccgggggtgt gagactcatg ccttcactgc
     5341 tcacccagag ggcaaacacc actccctgtc cttttcagtt taaacgaatt cgggctcggt
     5401 acccggggat cctctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag
     5461 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5521 gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     5581 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     5641 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     5701 gttcaggggg agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa
     5761 atcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     5821 catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa
     5881 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     5941 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6001 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6061 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     6121 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6181 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6241 gcttttttgg aggcctaggc ttttgcaaaa agcttaatta actgttgaca attaatcatc
     6301 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga
     6361 tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     6421 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     6481 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6541 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6601 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6661 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6721 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6781 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6841 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6901 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6961 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     7021 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     7081 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     7141 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     7201 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     7261 ttttgtgtga atcgatagcg ataaggatcc gctcatgcga tgataagctg tcaaacatga
     7321 gaattacaac ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg
     7381 cactgaaatc tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg
     7441 tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa
     7501 ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc
     7561 aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa
     7621 atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa
     7681 gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt
     7741 ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc
     7801 ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg
     7861 gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga
     7921 tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt
     7981 gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc
     8041 cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga
     8101 gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt
     8161 ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag
     8221 tatacactcc gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca
     8281 tgagattacg cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt
     8341 ctgccctcgt agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga
     8401 ggatcgagat ct
//
