LOCUS       sequence-317039-        7410 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses constitutively active AMPKalpha2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7410)
  TITLE     Birnbaum lab plasmids
  JOURNAL   Unpublished
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: 
            "Unpublished"
COMMENT     Sequence Label: pAMPK alpha2 delta312X
FEATURES             Location/Qualifiers
     source          1..7410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             716..745
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(2729..2747)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2730..2747)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2767..2784)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2773..2997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3043..3471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3130..3149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3340..3361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3480..3500)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3485..3814
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3665..3800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3727..3746
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3881..4675
                     /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(3935..3954)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4545..4564
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4849..4970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4886..4905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4940..4959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5019..5035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5019..5035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5032..5054)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5043..5059
                     /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(5067..5073,5074..5091,5092..5097))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5112..5133
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5250..5267)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5421..6009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5501..5520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6180..6971,6972..7040))
                     /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     6803..6822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7041..7145)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7220..7239)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccatggagca
      721 gaagcttatc tccgaggagg acctcggtgg cggcgagaag cagaagcacg acgggcgtgt
      781 gaagatcgga cactacgtgc tgggggacac cctgggcgtc ggcaccttcg gcaaagtgaa
      841 gattggagaa catcaattga caggccataa agtggcagtt aagatcttaa atagacagaa
      901 gattcgcagt ttagatgttg ttggaaaaat aaaacgagaa attcaaaatc ttaaactctt
      961 tcgtcatcct catattatca aactctacca agtgatcagc actccaacag acttttttat
     1021 ggtaatggaa tatgtgtctg gaggtgaatt gttcgactac atctgtaaac acgggagggt
     1081 tgaagaggtg gaagctcgcc ggctcttcca gcagattctg tctgccgtgg actactgtca
     1141 caggcacatg gttgtccaca gggacctgaa gccagagaac gtgttgctgg acgcccagat
     1201 gaatgctaag atagctgact tcggactctc taatatgatg tcagatggtg aatttctacg
     1261 aactagctgt ggatcgccaa attatgcagc accggaggtc atctcaggaa ggctgtatgc
     1321 gggtcctgag gttgatatct ggagctgtgg tgttatcctg tatgcccttc tctgtggcac
     1381 cctcccgttc gacgatgagc acgtgcctac gctctttaag aagatccgag ggggtgtgtt
     1441 ctacatcccg gagtatctca accgttctat tgccactctg ctgatgcaca tgctgcaggt
     1501 ggaccccttg aagcgagcaa ctatcaaaga catacgagag catgaatggt ttaaacagga
     1561 tttgcccagt tacctctttc ctgaagaccc ctcctatgat gctaacgtca ttgatgatga
     1621 ggctgtgaaa gaagtatgtg aaaaatttga gtgtacagaa tcagaagtga tgaacagttt
     1681 atagagtggt gaccctcaag accagctcgc agtggcttat catctcatca ttgacaatcg
     1741 gagaataatg aaccaagcca gtgagttcta cctcgcctcc agtcctccaa cgggttcctt
     1801 catggacgat agtgccatgc atattccccc cggcctgaaa ccacatcctg aaaggatgcc
     1861 acctctcata gcagacagcc ccaaagcacg ctgtccactg gatgcactca acacaactaa
     1921 gcccaaatct ttagctgtga aaaaagccaa gtggcacctt gggatccgaa gccagagcaa
     1981 accatacgac attatggcgg aggtgtaccg agctatgaag cagctggact ttgaatggaa
     2041 ggtagtgaat gcataccatc ttcgagtaag aagaaaaaac ccagtgactg gcaattacgt
     2101 gaaaatgagc ttacagcttt acctggttga caatcggagc tatcttctag actttaaaag
     2161 catcgatgat gaggtggtgg agcagaggtc tggttcttca acacctcagc gctcctgttc
     2221 tgctgccggc ctccacagac ctcggtcaag tgtcgattcc agcacagccg agaaccattc
     2281 actgtctggc tctctcactg gttctttgac tggcagcact ttgtcctccg cttccccgcg
     2341 cctgggcagt cataccatgg atttttttga aatgtgcgcc agtcttatca ctgctttagc
     2401 ccgttgataa cccaccaccg gtctctgtct ttctgttacc gcactgtgaa atcacataca
     2461 ctcttcaaat tattaccgca ctctcgggta ccacaggctc tgcaatagaa gttatgtgaa
     2521 cattcccagg tgacatgcag tgctgctgga aacacagaaa tctggctttc tgtttacttt
     2581 tagaactctg taactctgct gtgcctatga taggtatcaa tagctaggaa cggctgagtg
     2641 ctggtgaagc ttgttaactt acacccgtga attctgcaga tatccatcac actggcggcc
     2701 gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     2761 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2821 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2881 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2941 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3001 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3061 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3121 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3181 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3241 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3301 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3361 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3421 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3481 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3541 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3601 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3661 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3721 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3781 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3841 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3901 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3961 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4021 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4081 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4141 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4201 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4261 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4321 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4381 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4441 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4501 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4561 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4621 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4681 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4741 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4801 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4861 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4921 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4981 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5041 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5101 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5161 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5221 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5281 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5341 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5401 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5461 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5521 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5581 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5641 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5701 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5761 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5821 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5881 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5941 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6001 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6061 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6121 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6181 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6241 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6301 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6361 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6421 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6481 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6541 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6601 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6661 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6721 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6781 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6841 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6901 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6961 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7021 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7081 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7141 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     7201 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     7261 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     7321 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     7381 ttcgcgatgt acgggccaga tatacgcgtt
//
