LOCUS       sequence-473360-        6145 bp    DNA     circular SYN 14-JUL-2025
DEFINITION  Express HaloTag(R)-PP1Cg 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 6145)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6145)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HaloTag-PP1Cg Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6145
                     /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(3088..3106)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3143..3264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3154..3173)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3208..3227
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3509..3866
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3509..3700
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3532..3552)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3717..3852
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3881..3928
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3960..4754
                     /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(4014..4033)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4624..4643
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4818..4866
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5105..5693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5185..5204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5809..6092
                     /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 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 ggatttagat aaactcaaca tcgacagcat
     2041 tatccaacgg ctgctggaag tgagagggtc caagcctggt aagaatgtcc agcttcagga
     2101 gaatgaaatc agaggactgt gcttaaagtc tcgtgaaatc tttctcagtc agcctatcct
     2161 actagaactt gaagcaccac tcaaaatatg tggtgacatc catggacaat actatgattt
     2221 gctgcgactt tttgagtacg gtggtttccc accagaaagc aactacctgt ttcttgggga
     2281 ctatgtggac aggggaaagc agtcattgga gacgatctgc ctcttactgg cctacaaaat
     2341 aaaatatcct gagaattttt ttcttctcag agggaaccat gaatgtgcca gcatcaacag
     2401 aatttatgga ttttatgatg aatgtaaaag aagatacaac attaaactat ggaaaacttt
     2461 cacagactgt tttaactgtt taccgatagc agccatcgtg gatgagaaga tattctgctg
     2521 tcatggaggt ttatcaccag atcttcaatc tatggagcag attcggcgaa ttatgcgacc
     2581 aactgatgta ccagatcaag gtcttctttg tgatcttttg tggtctgacc ccgataaaga
     2641 tgtcttaggc tggggtgaaa atgacagagg agtgtccttc acatttggtg cagaagtggt
     2701 tgcaaaattt ctccataagc atgatttgga tcttatatgt agagcccatc aggtggttga
     2761 agatggatat gaattttttg caaagaggca gttggtcact ctgttttctg cgcccaatta
     2821 ttgcggagag tttgacaatg caggtgccat gatgagtgtg gatgaaacac taatgtgttc
     2881 ttttcagatt ttaaagcctg cagagaaaaa gaagccaaat gccacgagac ctgtaacgcc
     2941 tccaaggggt atgatcacaa agcaagcaaa gaaagtttaa acgaattcgg gctcggtacc
     3001 cggggatcct ctagagtcga cctgcaggca tgcaagctga tccggctgct aacaaagccc
     3061 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     3121 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3181 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3241 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3301 cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc
     3361 gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat
     3421 ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct
     3481 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
     3541 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3601 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3661 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3721 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3781 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3841 tttttggagg cctaggcttt tgcaaaaagc ttaattaact gttgacaatt aatcatcggc
     3901 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     3961 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4021 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4081 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4141 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4201 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4261 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4321 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4381 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4441 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4501 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4561 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4621 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4681 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4741 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4801 gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     4861 tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt gtccagatag
     4921 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacccgg
     4981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5041 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5281 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5521 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5641 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     5701 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5761 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaaatacag
     5821 gaacgcacgc tggatggccc ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag
     5881 tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt catacggtta
     5941 aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt tttacctgtc
     6001 tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg attatggtaa
     6061 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
     6121 cgtccggcgt agaggatcga gatct
//
