LOCUS       sequence-137644-        8406 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 8406)
  TITLE     Birnbaum lab plasmids
REFERENCE   2  (bases 1 to 8406)
  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"
COMMENT     Sequence Label: pMIGR AMPK KD
FEATURES             Location/Qualifiers
     source          1..8406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     332..354
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     370..386
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             439..468
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    2396..2963
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2413..2965
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2557..2574)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2784..2803
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2977..2979,2980..2982,2983..3696)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3022..3043)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3283..3302)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3630..3651
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3798..4312
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4474..4490
                     /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(4498..4504,4505..4522,4523..4528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4543..4564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4681..4698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4852..5440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4932..4951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5611..6402,6403..6471))
                     /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     6234..6253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6472..6576)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6644..6662
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6700..6722)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6822..6841
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7035..7057
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7185..7204
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7404..7920
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7984..8325
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             join(8392..8406,1..402)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag
       61 tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt
      121 aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc
      181 aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg
      241 gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg
      301 cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct
      361 cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag atctctcgag
      421 gttaacgaat tcgccatgga gcagaagctt atctccgagg aggacctcgg tggcggcgag
      481 aagcagaagc acgacgggcg tgtgaagatc ggacactacg tgctggggga caccctgggc
      541 gtcggcacct tcggcaaagt gaagattgga gaacatcaat tgacaggcca taaagtggca
      601 gttaggatct taaatagaca gaagattcgc agtttagatg ttgttggaaa aataaaacga
      661 gaaattcaaa atcttaaact ctttcgtcat cctcatatta tcaaactcta ccaagtgatc
      721 agcactccaa cagacttttt tatggtaatg gaatatgtgt ctggaggtga attgttcgac
      781 tacatctgta aacacgggag ggttgaagag gtggaagctc gccggctctt ccagcagatt
      841 ctgtctgccg tggactactg tcacaggcac atggttgtcc acagggacct gaagccagag
      901 aacgtgttgc tggacgccca gatgaatgct aagatagctg acttcggact ctctaatatg
      961 atgtcagatg gtgaatttct acgaactagc tgtggatcgc caaattatgc agcaccggag
     1021 gtcatctcag gaaggctgta tgcgggtcct gaggttgata tctggagctg tggtgttatc
     1081 ctgtatgccc ttctctgtgg caccctcccg ttcgacgatg agcacgtgcc tacgctcttt
     1141 aagaagatcc gagggggtgt gttctacatc ccggagtatc tcaaccgttc tattgccact
     1201 ctgctgatgc acatgctgca ggtggacccc ttgaagcgag caactatcaa agacatacga
     1261 gagcatgaat ggtttaaaca ggatttgccc agttacctct ttcctgaaga cccctcctat
     1321 gatgctaacg tcattgatga tgaggctgtg aaagaagtat gtgaaaaatt tgagtgtaca
     1381 gaatcagaag tgatgaacag tttatacagt ggtgaccctc aagaccagct cgcagtggct
     1441 tatcatctca tcattgacaa tcggagaata atgaaccaag ccagtgagtt ctacctcgcc
     1501 tccagtcctc caacgggttc cttcatggac gatagtgcca tgcatattcc ccccggcctg
     1561 aaaccacatc ctgaaaggat gccacctctc atagcagaca gccccaaagc acgctgtcca
     1621 ctggatgcac tcaacacaac taagcccaaa tctttagctg tgaaaaaagc caagtggcac
     1681 cttgggatcc gaagccagag caaaccatac gacattatgg cggaggtgta ccgagctatg
     1741 aagcagctgg actttgaatg gaaggtagtg aatgcatacc atcttcgagt aagaagaaaa
     1801 aacccagtga ctggcaatta cgtgaaaatg agcttacagc tttacctggt tgacaatcgg
     1861 agctatcttc tagactttaa aagcatcgat gatgaggtgg tggagcagag gtctggttct
     1921 tcaacacctc agcgctcctg ttctgctgcc ggcctccaca gacctcggtc aagtgtcgat
     1981 tccagcacag ccgagaacca ttcactgtct ggctctctca ctggttcttt gactggcagc
     2041 actttgtcct ccgcttcccc gcgcctgggc agtcatacca tggatttttt tgaaatgtgc
     2101 gccagtctta tcactgcttt agcccgttga taacccacca ccggtctctg tctttctgtt
     2161 accgcactgt gaaatcacat acactcttca aattattacc gcactctcgg gtaccacagg
     2221 ctctgcaata gaagttatgt gaacattccc aggtgacatg cagtgctgct ggaaacacag
     2281 aaatctggcc ttctgtttac ttttagaact ctgtaactct gctgtgccta tgataggtat
     2341 caatagctag gaacggctga gtgctggtga agcttgttaa cttacacccg tgaattccgc
     2401 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     2461 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     2521 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     2581 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     2641 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     2701 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     2761 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     2821 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     2881 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     2941 tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggagctg
     3001 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     3061 agcgtgtctg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     3121 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     3181 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     3241 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     3301 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     3361 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3421 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc
     3481 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3541 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3601 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3661 gggatcactc tcggcatgga cgagctgtac aagtaatgaa ttaattaaga attatcaagc
     3721 ttatcgatac cgtcgacctg cagccaagct tatcgataaa ataaaagatt ttatttagtc
     3781 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     3841 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     3901 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     3961 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     4021 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     4081 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     4141 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     4201 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     4261 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atgggtaaca
     4321 gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa gtaatcctga
     4381 ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag ttcattatgg
     4441 acagcgcaga aagagctggg gagaattgtg aaattgttat ccgctcacaa ttccacacaa
     4501 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4561 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5281 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5521 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5581 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5641 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5701 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5761 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5821 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5881 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5941 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6001 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6061 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6121 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6181 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6241 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6301 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6361 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6421 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6481 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6541 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6601 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6661 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6721 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6781 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6841 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6901 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6961 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7021 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc
     7081 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     7141 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     7201 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     7261 aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact
     7321 caacaatatc accagctgaa gcctatagag tacgagccat agataaaata aaagatttta
     7381 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     7441 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     7501 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     7561 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     7621 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     7681 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     7741 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     7801 cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc
     7861 gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt
     7921 tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc cccgccggga
     7981 ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc
     8041 atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
     8101 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
     8161 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
     8221 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
     8281 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
     8341 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
     8401 gttacc
//
