LOCUS       sequence-479770-        5065 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express H2A-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 5065)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5065)
  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            "Unpublished"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: H2A-HaloTag/CMV Vector
FEATURES             Location/Qualifiers
     source          1..5065
                     /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             1476..1496
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1512..2396
                     /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(2484..2502)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2539..2660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2550..2569)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2604..2623
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             3059..3853
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3113..3132)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3723..3742
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4025..4613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4105..4124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4729..5012
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 cgccatgtcc ggtcgtggta
     1081 agcagggtgg caaggcgcgc gccaaggcta agtcgcgctc gtcgcgcgcg gggctgcagt
     1141 tccccgtggg ccgcgtgcac cggttgctcc gcaagggcaa ctattcggag cgcgtgggcg
     1201 ccggcgcccc ggtctatctg gccgcggtgc tcgagtactt gactgccgag atcctggagc
     1261 ttgccggcaa cgcggcgcgc gacaacaaga agacgcgcat catcccgcgc cacctgcagc
     1321 tggccatccg caacgacgag gagctcaaca agctgctggg ccgcgtgacc atcgcgcagg
     1381 gtggcgtcct gcccaacatc caggccgtac tgctgcccaa gaagacggag agccaccaca
     1441 aggccaaggg caaggtttct ctcgagccaa ccactgagga tctgtacttt cagagcgata
     1501 acgatggatc cgaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg
     1561 gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc
     1621 acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc
     1681 atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt
     1741 atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag
     1801 aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca
     1861 atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg
     1921 acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc
     1981 gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc
     2041 gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
     2101 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
     2161 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
     2221 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
     2281 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
     2341 cggacctgat cggcagcgag atcgcgcgct ggctgtctac tctggagatt tccggttaat
     2401 agaattctag agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa
     2461 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc
     2521 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2581 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2641 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2701 gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat
     2761 tctaatggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     2821 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc
     2881 agcccttgcg ccctgagtgc ttgcggcagc gtgagcttca aaagaattgc cagctggggc
     2941 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag
     3001 gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgacggt cgtttcgcat
     3061 gcttgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3121 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3181 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3241 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3301 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3361 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3421 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3481 cgagcgagca cgcactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3541 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgta tgccggatgg
     3601 tgaggatctc gtcgtgactc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3661 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3721 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3781 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3841 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaaccgg
     3901 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3961 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4021 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4081 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4141 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4201 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4261 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4321 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4381 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4441 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4501 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4561 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     4621 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4681 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaaatacag
     4741 gaacgcacgc tggatggccc ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag
     4801 tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta
     4861 aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc
     4921 tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa
     4981 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
     5041 cgtccggcgt agaggatcga gatct
//
