LOCUS       sequence-501946-        6717 bp    DNA     circular SYN 04-MAR-2026
DEFINITION  Express AMPKa-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 6717)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6717)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: AMPKa-HaloTag Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1372..1392)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(1405..2295)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     regulatory      2292..2301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2310..2329)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2311..2329)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2371..2390)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2373..2505
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2641..2844)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2670..2690)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2845..3224
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3669..3686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3803..4348)
                     /direction=LEFT
                     /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     complement(3883..3902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4505..4553
                     /note="synthetic polyadenylation signal"
     CDS             complement(4617..5411)
                     /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(4728..4747)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5338..5357
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5443..5490)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5505..5862)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5519..5654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5671..5862
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5819..5839
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    6107..6228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6144..6163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6198..6217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6265..6283
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 tgtcacagga ttcttccttc gtacacgcaa ataatatggg tttacaacct tccattcata
       61 atccaattgt ttgattgctc tacatacttc tgccataata tcatttggtc gactttgact
      121 tctaattcct aaatgccatt ttgctttcct tacaccttgg tgtttggatt tctgtggatt
      181 taattcatca agggtatggc gtgcccttgg tgtttcagca accaagaatg gtactctttc
      241 aggatggggc cgagtcaggt gatgatcatc aagaaaagaa tcaggtgggc ttgtcgccaa
      301 atagaaatct ttggcttcat tcattattct cctgttatct attatgagat ggtaggcaac
      361 tgccaaagga tcctggtgat ttctgttgta aagacagctg agaacttcct cttctgagca
      421 ctcaaacttt tcacatactt cttttaaggc ttcatcatca atcatggttg aactatatga
      481 tggatcctca ggaaagagat attttggaag gtcctgtttg aaccattcat gttccctgat
      541 atctttgatt gtggccctct tcatgggatc cacctgcagc atatgtttca aaaggctaat
      601 cacagaagga tttaaatatt gaggggtata gaagatccca tcacatatct tcttaaaaag
      661 agttggcaca tggtcatcat caaatggaag ggttccacat aataaagcat agagaataac
      721 cccactgctc catatatcta cctctgggcc tgcatacaat cttcctgaaa ttacttctgg
      781 tgcagcatag ttgggtgagc cacaacttgt tcttaaaaat tcaccatctg acatcatgtt
      841 tgaaagacca aaatcagcta tctttgcatt catgtgtgca tcaagcagga cattttcagg
      901 tttcaaatct ctatggacca ccatatgcct gtgacaataa tccacaccag aaaggatctg
      961 ttggaacaga cgccgacttt ctttttcatc cagccttcca ttcttacaga tataatcaaa
     1021 tagctctcct cctgagacat attccatcac catgaaaata tcagatggtg tactgatgac
     1081 ctggtacagt ttaattatat gaggatgcct gaaaagcttg aggttctgaa tttctctgcg
     1141 gatttttcct accacatcaa ggctccgaat cttctgtcga ttgagtatct tcacagctac
     1201 tttatgccca gtcaattcat gtttgccaac cttcactttg ccgaaggtgc cgacccccag
     1261 cgtgtcaccc agaatgtagt ggccgatctt cacccgcccg tcgtgtttct gcttctcggc
     1321 tgtcgccatc tttctccagg aactgagtct gcgcatggcg atcgcgttat cgctctgaaa
     1381 gtacagatcc tcagtggttg gctcgccgga aatctcgagc gtcgacagcc agcgcgcgat
     1441 ctcgctgccg atcaggtccg ggttgtcttc ttgcagcaga ttcagacccg ggccgatgtc
     1501 cacagccttg cagttaggca ggcttttggc caggcgagcg gcttcggccg gtgggatcag
     1561 aacgcctggg gtgccccaga acagcagctt cgggacaggg gactggtgca gccagtccat
     1621 gtattcttcg accagcgcga cgatgttcgc tggctcaccg gcgattggca gctcgtttgg
     1681 gaagcgccac agtggctcgc ggtcaacagg attcaggaac ggctcgcggt aatggtccat
     1741 ctcgacttca gtcagcgggc ggacgacacc catcggcagc gtaccctcga taaaaacgtt
     1801 ctgatcgatg atcagcttgc ggccgacgtc ggtggtgcgg aaggcctgga aggtctcgcg
     1861 ggcaaattct ggccattcgt cccaggtcgg gatagggcgg atgaactcca taaatgcaat
     1921 acctttgacg cgctctggat tgcgcttggc ccagtggaaa cccagagcgg agccccagtc
     1981 gtgaatgacc aggacgacct cttccagacc cagggcttcg atgaaggcat ccatgaagcg
     2041 gacgtggtcg tcgaagaaat aacccaggtc tggtttgtcg gatttgccca taccgatcag
     2101 gtctggagca atgcagcgat gggtcggtgc aacatgcggg atgatgttgc gccacacgta
     2161 ggaggaggtc gggttaccgt gcaggaacag cacaggggtg ccatcgcgcg gaccaacatc
     2221 gacgtagtgc atgcgctcgc ccaggacttc cacataatgg gggtcgaatg gaaagccagt
     2281 accgatttct gccatggtgg ctttgctagc cctatagtga gtcgtattaa tactctagcc
     2341 ttaagagctg taattgaact gggagtggac acctgtggag agaaaggcaa agtggatgtc
     2401 agtaagacca ataggtgcct atcagaaacg caagagtctt ctctgtctcg acaagcccag
     2461 tttctattgg tctccttaaa cctgtcttgt aaccttgata cttacctgcc cagtgcctca
     2521 cgaccaactt ctgcagctta agttcgagac tgttgtgtca gaagcactga ctgcgttagc
     2581 aatttaactg tgataaacta ccgcaataaa gcttctagtg atctgacggt tcactaaacc
     2641 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaacggggc
     2701 ggggttatta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     2761 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     2821 ggtgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     2881 ccaagtagga aagtcccgta aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     2941 gtcattgacg tcaatagggg gcggacttgg catatgatac acttgatgta ctgccaagtg
     3001 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     3061 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     3121 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     3181 ccccgtaatt gattactatt aataactagt caataatcaa tgccaacatg gcggtcatat
     3241 tggacatgag ccaatataaa tgtacatatt atgatataga tacaacgtat gcaatggcca
     3301 atagccaata ttgatttatg ctatataacc aatgaataat atggctaatg gccaatattg
     3361 aagatctcga tcctctacgc cggacgcatc gtggccggag tactgcggtt tatctcgatg
     3421 gctacgaggg cagacagtaa gtggatttac cataatccct taattgtacg caccgctaaa
     3481 acgcgtaatc tcatgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct
     3541 agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca
     3601 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
     3661 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     3721 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     3781 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     3841 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3901 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     3961 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     4021 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     4081 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     4141 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     4201 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     4261 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     4321 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt
     4381 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
     4441 taagttgtaa ttctcatgtt tgacagctta tcatcgcatg agcggatcct tatcgctatc
     4501 gattcacaca aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca
     4561 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     4621 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     4681 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     4741 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     4801 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     4861 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     4921 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     4981 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     5041 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     5101 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5161 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5221 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5281 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5341 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5401 tgttcaatca tgatctgcct cctgggttta gttcctcacc ttgtcgtatt atactatgcc
     5461 gatatactat gccgatgatt aattgtcaac agttaattaa gctttttgca aaagcctagg
     5521 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     5581 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     5641 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     5701 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     5761 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     5821 cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc taaccaagtt
     5881 cctctttcag aggttatttc aggccatggt gctgcgcaga tccgcgtatg cggtgtgaaa
     5941 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     6001 gttaaaattc gattttacca catttgtaga ggttttactt gcttaaaaaa acctcccaca
     6061 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     6121 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6181 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
     6241 gaagcggccg ccccaagggg ttatgctagt tattgctcag cggtggcagc agccaactca
     6301 gcttcctttc gggctttgtt agcagccgga tcagcttgca tgcctgcagg tcgactctag
     6361 aggatccccg ggtaccgagc ccgaattcgt ttaaacttgt gcaagaattt taattagatt
     6421 tgcacacatc tcaaaaaatt ctattgtgtg acttccaggt cttggagtta ggtcaacagg
     6481 agaagagtca agtgaggtca cagatgaggt aagagaaact tctgaggatt ttccttgagc
     6541 ctcagcatct gaatcactcc tttggcaaga tcgatagttg ctaactgatc ccgatctctg
     6601 tggagtagca gtccctgatt tggcttctgt aatttcatca tcaatactac ggaaatccag
     6661 tagataagtt ctactatcca cttggtataa ctgtagactc attttggagt aagtgct
//
