LOCUS       sequence-473362-        6120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PP1Cg-HaloTag(R) 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 6120)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6120)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: PP1Cg-HaloTag Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6120
                     /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             2055..2075
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2091..2975
                     /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(3063..3081)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3118..3239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3129..3148)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3183..3202
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3484..3841
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3484..3675
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3507..3527)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3692..3827
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3856..3903
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3935..4729
                     /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(3989..4008)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4599..4618
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4793..4841
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5080..5668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5160..5179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5784..6067
                     /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 cgccatggcg gatttagata
     1081 aactcaacat cgacagcatt atccaacggc tgctggaagt gagagggtcc aagcctggta
     1141 agaatgtcca gcttcaggag aatgaaatca gaggactgtg cttaaagtct cgtgaaatct
     1201 ttctcagtca gcctatccta ctagaacttg aagcaccact caaaatatgt ggtgacatcc
     1261 atggacaata ctatgatttg ctgcgacttt ttgagtacgg tggtttccca ccagaaagca
     1321 actacctgtt tcttggggac tatgtggaca ggggaaagca gtcattggag acgatctgcc
     1381 tcttactggc ctacaaaata aaatatcctg agaatttttt tcttctcaga gggaaccatg
     1441 aatgtgccag catcaacaga atttatggat tttatgatga atgtaaaaga agatacaaca
     1501 ttaaactatg gaaaactttc acagactgtt ttaactgttt accgatagca gccatcgtgg
     1561 atgagaagat attctgctgt catggaggtt tatcaccaga tcttcaatct atggagcaga
     1621 ttcggcgaat tatgcgacca actgatgtac cagatcaagg tcttctttgt gatcttttgt
     1681 ggtctgaccc cgataaagat gtcttaggct ggggtgaaaa tgacagagga gtgtccttca
     1741 catttggtgc agaagtggtt gcaaaatttc tccataagca tgatttggat cttatatgta
     1801 gagcccatca ggtggttgaa gatggatatg aattttttgc aaagaggcag ttggtcactc
     1861 tgttttctgc gcccaattat tgcggagagt ttgacaatgc aggtgccatg atgagtgtgg
     1921 atgaaacact aatgtgttct tttcagattt taaagcctgc agagaaaaag aagccaaatg
     1981 ccacgagacc tgtaacgcct ccaaggggta tgatcacaaa gcaagcaaag aaagtttctc
     2041 tcgagccaac cactgaggat ctgtactttc agagcgataa cgatggatcc gaaatcggta
     2101 ctggctttcc attcgacccc cattatgtgg aagtcctggg cgagcgcatg cactacgtcg
     2161 atgttggtcc gcgcgatggc acccctgtgc tgttcctgca cggtaacccg acctcctcct
     2221 acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt gctccagacc
     2281 tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac gaccacgtcc
     2341 gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg
     2401 actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta
     2461 ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca gaatttgccc
     2521 gcgagacctt ccaggccttc cgcaccaccg acgtcggccg caagctgatc atcgatcaga
     2581 acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga
     2641 tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc
     2701 caaacgagct gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc gaagaataca
     2761 tggactggct gcaccagtcc cctgtcccga agctgctgtt ctggggcacc ccaggcgttc
     2821 tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg
     2881 acatcggccc gggtctgaat ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga
     2941 tcgcgcgctg gctgtctact ctggagattt ccggttaata gaattctaga gtcgacctgc
     3001 aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     3061 caccgctgag caataactag cataacccct tggggcggcc gcttcgagca gacatgataa
     3121 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3181 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3241 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     3301 taagcaagta aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt
     3361 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3421 gcggatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac
     3481 cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3541 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3601 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3661 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3721 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3781 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3841 aaagcttaat taactgttga caattaatca tcggcatagt atatcggcat agtataatac
     3901 gacaaggtga ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc
     3961 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4021 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4081 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4141 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4201 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4261 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4321 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4381 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4441 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4501 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4561 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4621 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4681 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4741 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat
     4801 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat
     4861 cctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     4921 tcagcaccgt ttctgcggac tggctttcta cccggtatca gctcactcaa aggcggtaat
     4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5521 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5641 gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga
     5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5761 cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc
     5821 tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc
     5881 tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca
     5941 gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg
     6001 ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag
     6061 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
