LOCUS       sequence-228263-        7410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7410)
  AUTHORS   Mu J, Brozinick JT, Valladares O, Bucan M, Birnbaum MJ
  TITLE     A role for AMP-activated protein kinase in contraction- and 
            hypoxia-regulated glucose transport in skeletal muscle.
  JOURNAL   Mol Cell. 2001 May . 7(5):1085-94.
  PUBMED    11389854
REFERENCE   2  (bases 1 to 7410)
  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: "Mol Cell. 
            2001 May . 7(5):1085-94."
COMMENT     Sequence Label: pAMPK alpha2 WT
FEATURES             Location/Qualifiers
     source          1..7410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..61)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     861..880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        861..879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             948..977
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2961..2979)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2962..2979)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2999..3016)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3005..3229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3275..3703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3362..3381)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3572..3593
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3712..3732)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3717..4046
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3897..4032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3959..3978
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4113..4907
                     /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(4167..4186)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4777..4796
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5081..5202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5118..5137)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5172..5191
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5251..5267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5251..5267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5264..5286)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5275..5291
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5299..5305,5306..5323,5324..5329))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5344..5365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5482..5499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5653..6241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5733..5752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6412..7203,7204..7272))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7035..7054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7273..7377)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
       61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
      181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga
      901 gctcggatcc actagtaacg gccgccagtg tgctggaatt cgccatggag cagaagctta
      961 tctccgagga ggacctcggt ggcggcgaga agcagaagca cgacgggcgt gtgaagatcg
     1021 gacactacgt gctgggggac accctgggcg tcggcacctt cggcaaagtg aagattggag
     1081 aacatcaatt gacaggccat aaagtggcag ttaagatctt aaatagacag aagattcgca
     1141 gtttagatgt tgttggaaaa ataaaacgag aaattcaaaa tcttaaactc tttcgtcatc
     1201 ctcatattat caaactctac caagtgatca gcactccaac agactttttt atggtaatgg
     1261 aatatgtgtc tggaggtgaa ttgttcgact acatctgtaa acacgggagg gttgaagagg
     1321 tggaagctcg ccggctcttc cagcagattc tgtctgccgt ggactactgt cacaggcaca
     1381 tggttgtcca cagggacctg aagccagaga acgtgttgct ggacgcccag atgaatgcta
     1441 agatagctga cttcggactc tctaatatga tgtcagatgg tgaatttcta cgaactagct
     1501 gtggatcgcc aaattatgca gcaccggagg tcatctcagg aaggctgtat gcgggtcctg
     1561 aggttgatat ctggagctgt ggtgttatcc tgtatgccct tctctgtggc accctcccgt
     1621 tcgacgatga gcacgtgcct acgctcttta agaagatccg agggggtgtg ttctacatcc
     1681 cggagtatct caaccgttct attgccactc tgctgatgca catgctgcag gtggacccct
     1741 tgaagcgagc aactatcaaa gacatacgag agcatgaatg gtttaaacag gatttgccca
     1801 gttacctctt tcctgaagac ccctcctatg atgctaacgt cattgatgat gaggctgtga
     1861 aagaagtatg tgaaaaattt gagtgtacag aatcagaagt gatgaacagt ttatacagtg
     1921 gtgaccctca agaccagctc gcagtggctt atcatctcat cattgacaat cggagaataa
     1981 tgaaccaagc cagtgagttc tacctcgcct ccagtcctcc aacgggttcc ttcatggacg
     2041 atagtgccat gcatattccc cccggcctga aaccacatcc tgaaaggatg ccacctctca
     2101 tagcagacag ccccaaagca cgctgtccac tggatgcact caacacaact aagcccaaat
     2161 ctttagctgt gaaaaaagcc aagtggcacc ttgggatccg aagccagagc aaaccatacg
     2221 acattatggc ggaggtgtac cgagctatga agcagctgga ctttgaatgg aaggtagtga
     2281 atgcatacca tcttcgagta agaagaaaaa acccagtgac tggcaattac gtgaaaatga
     2341 gcttacagct ttacctggtt gacaatcgga gctatcttct agactttaaa agcatcgatg
     2401 atgaggtggt ggagcagagg tctggttctt caacacctca gcgctcctgt tctgctgccg
     2461 gcctccacag acctcggtca agtgtcgatt ccagcacagc cgagaaccat tcactgtctg
     2521 gctctctcac tggttctttg actggcagca ctttgtcctc cgcttccccg cgcctgggca
     2581 gtcataccat ggattttttt gaaatgtgcg ccagtcttat cactgcttta gcccgttgat
     2641 aacccaccac cggtctctgt ctttctgtta ccgcactgtg aaatcacata cactcttcaa
     2701 attattaccg cactctcggg taccacaggc tctgcaatag aagttatgtg aacattccca
     2761 ggtgacatgc agtgctgctg gaaacacaga aatctggcct tctgtttact tttagaactc
     2821 tgtaactctg ctgtgcctat gataggtatc aatagctagg aacggctgag tgctggtgaa
     2881 gcttgttaac ttacacccgt gaattctgca gatatccatc acactggcgg ccgctcgagc
     2941 atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     3001 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3061 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3121 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3181 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3241 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3301 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3361 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3421 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3481 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3541 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3601 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3661 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3721 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3781 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3841 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3901 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3961 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4021 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4081 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4141 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4201 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4261 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4321 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4381 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4441 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4501 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4561 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4621 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4681 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4741 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4801 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4861 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4921 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4981 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5041 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5101 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5161 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5221 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5281 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5341 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5401 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5461 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5521 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5581 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6061 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6121 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6181 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6241 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6301 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6361 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6421 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6481 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6541 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6601 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6661 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6721 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6781 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6841 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6901 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6961 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7021 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7081 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7141 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7201 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7261 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7321 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7381 atttccccga aaagtgccac ctgacgtcga
//
