LOCUS       sequence-452452-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MYC-tagged hTRAK1 (S200A, S201A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  TITLE     Energy stress activates AMPK to arrest mitochondria via 
            phosphorylation of TRAK1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7146)
  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            "Unpublished"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: MYC-hTRAK1(S200A, S201A)
FEATURES             Location/Qualifiers
     source          1..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1317..1335
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      1367..1376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1376..1405
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4344..4363)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4345..4363)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4637..4758
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4674..4693)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4728..4747
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4765..5220)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4902..4923)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5114..5133
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5247..5351
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5353..5710
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5353..5544
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5376..5396)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5561..5696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5745..6539
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5745..6539
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5799..5818)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6409..6428
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6727..6746)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6771..6818
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg attacgccaa gctcgaaatt
     1321 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ccatggagca
     1381 gaaactcatc tctgaagagg atctggcccg ggcggatcca atggcattgg tttttcaatt
     1441 cgggcagccc gtcagggctc agcctctgcc aggactctgc cacggcaagc tcattcggac
     1501 aaacgcctgt gatgtgtgca acagcaccga tcttccggaa gtcgagatca ttagcctgct
     1561 ggaggagcag ctgccccatt ataagttaag agccgacacc atctacggtt atgaccacga
     1621 cgactggctc catacacctc tcatttctcc agatgccaac attgacctca caaccgagca
     1681 aattgaagag acgttaaaat acttcctttt atgtgctgaa agagttggcc agatgactaa
     1741 gacatataat gacatagatg ctgtcactcg gcttcttgag gagaaagagc gggatttaga
     1801 attggccgct cgcatcggcc agtcgttgtt gaagaagaac aagaccctaa ccgagaggaa
     1861 cgagctgctg gaggagcagg tggaacacat cagggaggag gtgtctcagc tccggcatga
     1921 gctgtccatg aaggatgagc tgcttcagtt ctacaccagc gctgcggagg agagtgagcc
     1981 cgagtccgtt tgctcaaccc cgttgaagag gaatgaggcg gcctcctcag tccagaatta
     2041 ctttcatttg gattctcttc aaaagaagct gaaagacctt gaagaggaga atgttgtact
     2101 tcgatccgag gccagccagc tgaagacaga gaccatcacc tatgaggaga aggagcagca
     2161 gctggtcaat gactgcgtga aggagctgag ggatgccaat gtccagattg ctagtatctc
     2221 agaggaactg gccaagaaga cggaagatgc tgcccgccag caagaggaga tcacacacct
     2281 gctatcgcaa atagttgatt tgcagaaaaa ggcaaaagct tgcgcagtgg aaaatgaaga
     2341 acttgtccag catctggggg ctgctaagga tgcccagcgg cagctcacag ccgagctgcg
     2401 tgagctggag gacaagtacg cagagtgcat ggagatgctg catgaggcgc aggaggagct
     2461 gaagaacctc cggaacaaaa ccatgcccaa taccacgtct cggcgctacc actcgctggg
     2521 cctgtttccc atggattcct tggcagcaga gattgaggga acgatgcgca aggagctgca
     2581 gttggaagag gccgagtctc cagacatcac tcaccagaag cgtgtctttg agacagtaag
     2641 aaacatcaac caggttgtca agcagagatc tctgacccct tctcccatga acatccccgg
     2701 ctccaaccag tcctcggcca tgaactccct cctgtccagc tgcgtcagca ccccccggtc
     2761 cagcttctac ggcagcgaca taggcaacgt cgtcctcgac aacaagacca acagcatcat
     2821 tctggaaaca gaggcagccg acctgggaaa cgatgagcgg agtaagaagc cggggacgcc
     2881 gggcacccca ggctcccacg acctggagac ggcgctgagg cggctgtccc tgcgccggga
     2941 gaactacctc tcggagagga ggttctttga ggaggagcaa gagaggaagc tccaggagct
     3001 ggcggagaag ggcgagctgc gcagcggctc cctcacaccc actgagagca tcatgtccct
     3061 gggcacgcac tcccgcttct ccgagttcac cggcttctct ggcatgtcct tcagcagccg
     3121 ctcctacctg cctgagaagc tccagatcgt gaagccgctg gaaggttctg ccacacttca
     3181 ccactggcag cagttggccc aacctcacct tgggggcatc ctggaccccc ggcctggtgt
     3241 ggtcaccaag ggcttccgga cgctggatgt tgacctggac gaagtgtact gccttaacga
     3301 ctttgaagaa gatgacacag gtgaccacat ttctctccca cgcctagcta cctccactcc
     3361 agttcagcac ccagagacct cagcgcacca tcctgggaag tgcatgtctc agaccaactc
     3421 caccttcacc ttcaccacct gtcgcatcct gcatccttca gatgagctca ctcgggtcac
     3481 accaagcctt aactcagccc caactccagc ttgtggcagc accagccact tgaaatccac
     3541 gccggtggcc acaccatgca ctccacggag actgagcctg gctgagtcct tcactaacac
     3601 ccgtgagtcc acgaccacca tgagcacatc cctggggctc gtgtggctgt tgaaggagcg
     3661 gggcatttct gctgccgtgt acgaccccca gagctgggac agggccggcc ggggctccct
     3721 cctgcactcc tacacgccca agatggctgt gatcccctct actccgccga actcgcctat
     3781 gcagacaccc acatcctccc caccctcctt tgagttcaag tgcacgagcc ctccctacga
     3841 caatttcctg gcttccaagc cagccagctc catcctgagg gaagtgagag aaaagaacgt
     3901 ccgcagcagc gagagccaga ccgacgtgtc cgtctccaac ctcaacctcg tggacaaagt
     3961 caggaggttt ggggtggcca aagtggtgaa ctcagggcga gcccatgtcc ccaccttgac
     4021 tgaggagcag ggacccctcc tctgtgggcc cccggggcca gcaccagccc ttgttcccag
     4081 aggcctggta cctgagggcc tgcccctcag atgccccact gtcaccagtg ccatcggtgg
     4141 gctgcagctc aatagtggca tccggcggaa tcgcagcttc cccaccatgg tgggatctag
     4201 catgcagatg aaagctcctg tgactctcac ctcgggcatc ttgatgggtg ctaagctctc
     4261 caaacaaact agcttacggt aagtctcgag ggggggcccg gtaccttaat taattaaggt
     4321 accaggtaag tgtacccaat tcgccctata gtgagtcgta ttacaattca ctcgatcgcc
     4381 cttcccaaca gttgcgcagc ctgaatggcg aatggagatc caatttttaa gtgtataatg
     4441 tgttaaacta ctgattctaa ttgtttgtgt attttagatt cacagtccca aggctcattt
     4501 caggcccctc agtcctcaca gtctgttcat gatcataatc agccatacca catttgtaga
     4561 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4621 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4681 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4741 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     4801 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4861 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4921 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4981 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5041 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5101 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5161 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5221 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5281 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5341 aaaggaagaa tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     5401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5461 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5521 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5581 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5641 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5701 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5761 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5821 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5881 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5941 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6001 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6061 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6121 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6181 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagaacat caggggctcg
     6241 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     6301 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6361 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6421 gtgatattgc tgaagaactt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6481 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6541 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6601 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6661 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     6721 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     6781 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6841 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6901 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6961 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7021 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7081 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7141 atcttc
//
