LOCUS       sequence-473340-        6790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HaloTag(R)-FYN 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 6790)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6790)
  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            "Unpublished"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: HaloTag-FYN Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    3..84
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        317..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          960..1092
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1075..1094
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1136..1155
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1136..1154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1164..1173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1170..2060
                     /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             2073..2093
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     polyA_signal    3832..3953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3843..3862)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4068..4101)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4111..4158
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4190..4984
                     /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(4244..4263)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4854..4873
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5142..5730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5222..5241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5856..5904
                     /note="synthetic polyadenylation signal"
     CDS             complement(5928..6326)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6323..6332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6357..6714)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6371..6506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6523..6714
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6671..6691
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 cggcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
       61 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      121 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
      181 gattatgatc cgcggccgca atacgtcgac gttatcagct gacttcgtac gagagcctag
      241 gattatggcg cgccactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      841 agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      901 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg
      961 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1021 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1081 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtattaata
     1141 cgactcacta tagggctagc aaagccacca tggcagaaat cggtactggc tttccattcg
     1201 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1261 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     1321 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     1381 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     1441 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     1501 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     1561 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1621 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1681 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1741 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1801 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1861 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1921 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1981 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     2041 cgacgctcga gatttccggc gagccaacca ctgaggatct gtactttcag agcgataacg
     2101 cgatcgccat gggctgtgtg caatgtaagg ataaagaagc aacaaaactg acggaggaga
     2161 gggacggcag cctgaaccag agctctgggt accgctatgg cacagacccc acccctcagc
     2221 actaccccag cttcggtgtg acctccatcc ccaactacaa caacttccac gcagccgggg
     2281 gccaaggact caccgtcttt ggaggtgtga actcttcgtc tcatacgggg accttgcgta
     2341 cgagaggagg aacaggagtg acactctttg tggcccttta tgactatgaa gcacggacag
     2401 aagatgacct gagttttcac aaaggagaaa aatttcaaat attgaacagc tcggaaggag
     2461 attggtggga agcccgctcc ttgacaactg gagagacagg ttacattccc agcaattatg
     2521 tggctccagt tgactctatc caggcagaag agtggtactt tggaaaactt ggccgaaaag
     2581 atgctgagcg acagctattg tcctttggaa acccaagagg tacctttctt atccgcgaga
     2641 gtgaaaccac caaaggtgcc tattcacttt ctatccgtga ttgggatgat atgaaaggag
     2701 accatgtcaa acattataaa attcgcaaac ttgacaatgg tggatactac attaccaccc
     2761 gggcccagtt tgaaacactt cagcagcttg tacaacatta ctcagagaga gctgcaggtc
     2821 tctgctgccg cctagtagtt ccctgtcaca aagggatgcc aaggcttacc gatctgtctg
     2881 tcaaaaccaa agatgtctgg gaaatccctc gagaatccct gcagttgatc aagagactgg
     2941 gaaatgggca gtttggggaa gtatggatgg gtacctggaa tggaaacaca aaagtagcca
     3001 taaagactct taaaccaggc acaatgtccc ccgaatcatt ccttgaggaa gcgcagatca
     3061 tgaagaagct gaagcacgac aagctggtcc agctctatgc agtggtgtct gaggagccca
     3121 tctacatcgt caccgagtat atgaacaaag gaagtttact ggatttctta aaagatggag
     3181 aaggaagagc tctgaaatta ccaaatcttg tggacatggc agcacaggtg gctgcaggaa
     3241 tggcttacat cgagcgcatg aattatatcc atagagatct gcgatcagca aacattctag
     3301 tggggaatgg actcatatgc aagattgctg acttcggatt ggcccgattg atagaagaca
     3361 atgagtacac agcaagacaa ggtgcaaagt tccccatcaa gtggacggcc cccgaggcag
     3421 ccctgtacgg gaggttcaca atcaagtctg acgtgtggtc ttttggaatc ttactcacag
     3481 agctggtcac caaaggaaga gtgccatacc caggcatgaa caaccgggag gtgctggagc
     3541 aggtggagcg aggctacagg atgccctgcc cgcaggactg ccccatctct ctgcatgagc
     3601 tcatgatcca ctgctggaaa aaggaccctg aagaacgccc cacttttgag tacttgcaga
     3661 gcttcctgga agactacttt accgcgacag agccccagta ccaacctggt gaaaacctgg
     3721 tttaaacgaa ttcgggctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
     3781 gctgatttgg ctgctgccac cgctgagcaa ggccgcttcg agcagacatg ataagataca
     3841 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3901 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3961 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     4021 agtaaaacct ctacaaatgt ggtaaaatct taattaagaa ttctaatata acttcgtata
     4081 gcatacatta tacgaagtta tggatccact gttgacaatt aatcatcggc atagtatatc
     4141 ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca
     4201 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4261 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4321 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4381 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4441 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4501 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4561 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4621 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4681 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4741 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4801 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4861 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4921 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4981 ctgattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     5041 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5101 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5161 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5221 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5281 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5341 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5401 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5461 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5521 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5581 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5641 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5701 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     5761 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5821 atcttcacct agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg
     5881 taatgaaaat aaagatattt tattgcggcc atcgtgatgg ctagagttta gccctcccac
     5941 acataaccag agggcagcaa ttcacgaatc ccaactgccg tcggctgtcc atcactgtcc
     6001 ttcactatgg ctttgatccc aggatgcaga tcgagaagca cctgtcggca ccgtccgcag
     6061 gggctcaaga tgcccctgtt ctcatttccg atcgcgacga tacaagtcag gttgccagct
     6121 gccgcagcag cagcagtgcc cagcaccacg agttctgcac aaggtccccc agtaaaatga
     6181 tatacattga caccagtgaa gatgcggccg tcgctagaga gagctgcgct ggcgacgctg
     6241 tagtcttcag agatggggat gctgttgatt gtagccgttg ctctttcaat gagggtggat
     6301 tcttcttgag acaaaggctt ggccatggtg gcgctagtgt cagaagaatc gagctttttg
     6361 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     6421 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     6481 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     6541 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     6601 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     6661 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta
     6721 cctaaccaag ttcctctttc agaggttatt tcaggccatg gtgctgcgca agacgcgtta
     6781 attaagacgt
//
