LOCUS       sequence-479801-        8387 bp    DNA     circular SYN 01-AUG-2025
DEFINITION  Express JAK3-HaloTag 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 8387)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8387)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: JAK3-HaloTag Vector
FEATURES             Location/Qualifiers
     source          1..8387
                     /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"
     CDS             4458..4478
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             4494..5378
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(5466..5484)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5521..5642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5532..5551)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5586..5605
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5887..6244
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5887..6078
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5910..5930)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6095..6230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6259..6306
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6338..7132
                     /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(6392..6411)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7002..7021
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7196..7244
                     /note="synthetic polyadenylation signal"
     rep_origin      7401..7946
                     /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     7847..7866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8063..8080
                     /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 ggctagcgat cgccatggca cctccaagtg
     1081 aagagacgcc cctgatccct cagcgttcat gcagcctctt gtccacggag gctggtgccc
     1141 tgcatgtgct gctgcccgct cggggccccg ggccccccca gcgcctatct ttctcctttg
     1201 gggaccactt ggctgaggac ctgtgcgtgc aggctgccaa ggccagcggc atcctgcctg
     1261 tgtaccactc cctctttgct ctggccacgg aggacctgtc ctgctggttc cccccgagcc
     1321 acatcttctc cgtggaggat gccagcaccc aagtcctgct gtacaggatt cgcttttact
     1381 tccccaattg gtttgggctg gagaagtgcc accgcttcgg gctacgcaag gatttggcca
     1441 gtgctatcct tgacctgcca gtcctggagc acctctttgc ccagcaccgc agtgacctgg
     1501 tgagtgggcg cctccccgtg ggcctcagtc tcaaggagca gggtgagtgt ctcagcctgg
     1561 ccgtgttgga cctggcccgg atggcgcgag agcaggccca gcggccggga gagctgctga
     1621 agactgtcag ctacaaggcc tgcctacccc caagcctgcg cgacctgatc cagggcctga
     1681 gcttcgtgac gcggaggcgt attcggagga cggtgcgcag agccctgcgc cgcgtggccg
     1741 cctgccaggc agaccggcac tcgctcatgg ccaagtacat catggacctg gagcggctgg
     1801 atccagccgg ggccgccgag accttccacg tgggcctccc tggggccctt ggtggccacg
     1861 acgggctggg gctgctccgc gtggctggtg acggcggcat cgcctggacc cagggagaac
     1921 aggaggtcct ccagcccttc tgcgactttc cagaaatcgt agacattagc atcaagcagg
     1981 ccccgcgcgt tggcccggcc ggagagcacc gcctggtcac tgttaccagg acagacaacc
     2041 agattttaga ggccgagttc ccagggctgc ccgaggctct gtcgttcgtg gcgctcgtgg
     2101 acggctactt ccggctgacc acggactccc agcacttctt ctgcaaggag gtggcaccgc
     2161 cgaggctgct ggaggaagtg gccgagcagt gccacggccc catcactctg gactttgcca
     2221 tcaacaagct caagactggg ggctcacgtc ctggctccta tgttctccgc cgcagccccc
     2281 aggactttga cagcttcctc ctcactgtct gtgtccagaa cccccttggt cctgattata
     2341 agggctgcct catccggcgc agccccacag gaaccttcct tctggttggc ctcagccgac
     2401 cccacagcag tcttcgagag ctcctggcaa cctgctggga tggggggctg cacgtagatg
     2461 gggtggcagt gaccctcact tcctgctgta tccccagacc caaagaaaag tccaacctga
     2521 tcgtggtcca gagaggtcac agcccaccca catcatcctt ggttcagccc caatcccaat
     2581 accagctgag tcagatgaca tttcacaaga tccctgctga cagcctggag tggcatgaga
     2641 acctgggcca tgggtccttc accaagattt accggggctg tcgccatgag gtggtggatg
     2701 gggaggcccg aaagacagag gtgctgctga aggtcatgga tgccaagcac aagaactgca
     2761 tggagtcatt cctggaagca gcgagcttga tgagccaagt gtcgtaccgg catctcgtgc
     2821 tgctccacgg cgtgtgcatg gctggagaca gcaccatggt gcaggaattt gtacacctgg
     2881 gggccataga catgtatctg cgaaaacgtg gccacctggt gccagccagc tggaagctgc
     2941 aggtggtcaa acagctggcc tacgccctca actatctgga ggacaaaggc ctgccccatg
     3001 gcaatgtctc tgcccggaag gtgctcctgg ctcgggaggg ggctgatggg agcccgccct
     3061 tcatcaagct gagtgaccct ggggtcagcc ccgctgtgtt aagcctggag atgctcaccg
     3121 acaggatccc ctgggtggcc cccgagtgtc tccgggaggc gcagacactt agcttggaag
     3181 ctgacaagtg gggcttcggc gccacggtct gggaagtgtt tagtggcgtc accatgccca
     3241 tcagtgccct ggatcctgct aagaaactcc aattttatga ggaccggcag cagctgccgg
     3301 cccccaagtg gacagagctg gccctgctga ttcaacagtg catggcctat gagccggtcc
     3361 agaggccctc cttccgagcc gtcattcgtg acctcaatag cctcatctct tcagactatg
     3421 agctcctctc agaccccaca cctggtgccc tggcacctcg tgatgggctg tggaatggtg
     3481 cccagctcta tgcctgccaa gaccccacga tcttcgagga gagacacctc aagtacatct
     3541 cacagctggg caagggcaac tttggcagcg tggagctgtg ccgctatgac ccgctaggcg
     3601 acaatacagg tgccctggtg gccgtgaaac agctgcagca cagcgggcca gaccagcaga
     3661 gggactttca gcgggagatt cagatcctca aagcactgca cagtgatttc attgtcaagt
     3721 atcgtggtgt cagctatggc ccgggccgcc agagcctgcg gctggtcatg gagtacctgc
     3781 ccagcggctg cttgcgcgac ttcctgcagc ggcaccgcgc gcgcctcgat gccagccgcc
     3841 tccttctcta ttcctcgcag atctgcaagg gcatggagta cctgggctcc cgccgctgcg
     3901 tgcaccgcga cctggccgcc cgaaacatcc tcgtggagag cgaggcacac gtcaagatcg
     3961 ctgacttcgg cctagctaag ctgctgccgc ttgacaaaga ctactacgtg gtccgcgagc
     4021 caggccagag ccccattttc tggtatgccc ccgaatccct ctcggacaac atcttctctc
     4081 gccagtcaga cgtctggagc ttcggggtcg tcctgtacga gctcttcacc tactgcgaca
     4141 aaagctgcag cccctcggcc gagttcctgc ggatgatggg atgtgagcgg gatgtccccg
     4201 ccctctgccg cctcttggaa ctgctggagg agggccagag gctgccggcg cctcctgcct
     4261 gccctgctga ggttcacgag ctcatgaagc tgtgctgggc ccctagccca caggaccggc
     4321 catcattcag cgccctgggc ccccagctgg acatgctgtg gagcggaagc cgggggtgtg
     4381 agactcatgc cttcactgct cacccagagg gcaaacacca ctccctgtcc ttttcagttt
     4441 ctctcgagcc aaccactgag gatctgtact ttcagagcga taacgatgga tccgaaatcg
     4501 gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc atgcactacg
     4561 tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac ccgacctcct
     4621 cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc attgctccag
     4681 acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc gacgaccacg
     4741 tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc ctggtcattc
     4801 acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag cgcgtcaaag
     4861 gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg ccagaatttg
     4921 cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc
     4981 agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg
     5041 agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca ctgtggcgct
     5101 tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat
     5161 acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc accccaggcg
     5221 ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg
     5281 tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg atcggcagcg
     5341 agatcgcgcg ctggctgtct actctggaga tttccggtta atagaattct agagtcgacc
     5401 tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     5461 tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga gcagacatga
     5521 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     5581 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     5641 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     5701 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
     5761 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5821 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     5881 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5941 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6061 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6121 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6241 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     6301 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
     6361 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     6421 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     6481 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     6541 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     6601 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6661 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     6721 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     6781 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6841 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     6901 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6961 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     7021 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     7081 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     7141 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     7201 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     7261 gatccgctca tgcgatgata agctgtcaaa catgagaatt acaacttata tcgtatgggg
     7321 ctgacttcag gtgctacatt tgaagagata aattgcactg aaatctagaa atattttatc
     7381 tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa
     7441 aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg
     7501 aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct
     7561 taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag
     7621 tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7681 gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc
     7741 ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg
     7801 taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga aacgcctggt
     7861 atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt tcgtgatgct
     7921 tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca cttccctgtt
     7981 aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg
     8041 ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat atatcctgta
     8101 tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg aagcacttca
     8161 ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag cgctgatgtc
     8221 cggcggtgct tttgccgtta cgcaccaccc cgtcatgaga ttacgcgttt tagcggtgcg
     8281 tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa
     8341 ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
