LOCUS       sequence-324232-        8004 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression vector for constitutively active AMPK alpha 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8004)
  AUTHORS   Liu X, Xiao ZD, Han L, Zhang J, Lee SW, Wang W, Lee H, Zhuang L, 
            Chen J, Lin HK, Wang J, Liang H, Gan B
  TITLE     LncRNA NBR2 engages a metabolic checkpoint by regulating AMPK under 
            energy stress.
  JOURNAL   Nat Cell Biol. 2016 Apr;18(4):431-42. doi: 10.1038/ncb3328. Epub 
            2016 Mar 21.
  PUBMED    26999735
REFERENCE   2  (bases 1 to 8004)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ncb3328"; 
            journalName: "Nat Cell Biol"; date: "2016-04"; volume: "18"; issue: 
            "4"; pages: "431-42"
COMMENT     Sequence Label: pLenti6-AMPK alpha1 (1-312)
FEATURES             Location/Qualifiers
     source          1..8004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     enhancer        1814..2117
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2118..2321
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2272..2292
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2318..2342
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2438..2462
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3435..3459)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3512..3553
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3590..3610)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3595..3924
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3775..3910
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3972..4019
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4038..4436
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4523..4756
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4828..4949
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4865..4884)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4919..4938
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4989..5124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5144..5163)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5145..5163)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5173..5190)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5173..5189)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5182..5204)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5331..5786
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5418..5437)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5628..5649
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5812..5916
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5917..5985,5986..6777)
                     /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     complement(6135..6154)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6948..7536
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7437..7456
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7690..7707
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7824..7845
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7860..7865,7866..7883,7884..7890)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7898..7914
                     /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)."
     primer_bind     7903..7925
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7922..7938
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7922..7938
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7957..7977
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7959..7977
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgataag
     1801 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     1861 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     1921 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     1981 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2041 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2101 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2161 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2221 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2281 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     2341 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagagga
     2401 tccactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca aaaaagcagg
     2461 ctccgcggcc gcccccttca ccatggccga gaagcagaag cacgacgggc gggtgaagat
     2521 cggccactac atcctggggg acacgctggg cgtcggcacc ttcgggaaag tgaaggtggg
     2581 caagcacgag ttgactggac ataaagttgc tgtgaagata ctcaaccggc agaagattcg
     2641 aagcctggac gtggtcggga aaatccgcag agagatccag aacctgaagc ttttcaggca
     2701 ccctcatata atcaaactgt accaggtcat cagtacaccg tctgatattt tcatggtcat
     2761 ggaatatgtc tcaggaggag agctatttga ttatatctgt aaaaatggaa ggttggacga
     2821 aaaggagagt cgacgtctgt tccagcagat cctttctggt gtggactatt gtcacaggca
     2881 tatggtggtc cacagagatt tgaaacctga aaacgtcctg cttgatgcac acatgaatgc
     2941 aaagatagcc gacttcggtc tttcaaacat gatgtcagat ggtgaatttt taagaacgag
     3001 ctgtggctcg cccaattatg ctgcaccaga agtaatttca ggaagattgt acgcaggccc
     3061 tgaagtagac atctggagca gcggggtcat tctctatgct ttgctgtgtg gaactctccc
     3121 ttttgatgat gaccacgtgc caactctttt taagaagata tgtgacggga tattttatac
     3181 ccctcagtat ttgaatccct ctgtaataag ccttttgaag catatgctgc aggtagatcc
     3241 tatgaagagg gccacaataa aagatatcag ggaacatgaa tggtttaagc aggaccttcc
     3301 aaaatatctc tttcctgaag acccgtctta tagttcaacc atgattgatg atgaagcctt
     3361 aaaagaagtg tgtgagaagt tcgagtgctc agaggaggag gtcctcagct gcctgtacaa
     3421 gggtgggcgc gccgacccag ctttcttgta caaagtggtt gatatccagc acagtggcgg
     3481 ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     3541 tctcgattct acgcgtaccg gttagtaatg agtttggaat taattctgtg gaatgtgtgt
     3601 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3661 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3721 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3781 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3841 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3901 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3961 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     4021 aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     4081 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     4141 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     4201 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     4261 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     4321 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     4381 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac
     4441 aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     4501 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     4561 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     4621 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4681 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4741 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     4801 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     4861 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4921 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     4981 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5041 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5101 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     5161 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5221 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5281 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     5341 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5401 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5461 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5521 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5581 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5641 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5701 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5761 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     5821 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5881 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5941 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6001 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6061 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6121 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6181 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6241 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6301 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6361 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6421 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6481 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6541 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6601 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6661 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6721 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6781 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6841 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7081 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7561 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7621 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7681 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7741 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7801 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7861 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7921 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     7981 caaaagctgg agctgcaagc ttaa
//
