LOCUS       sequence-202148-        7277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MBP-rPKL S113TAG for production and purification of rat PKL pS113.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7277)
  AUTHORS   Gassaway BM, Cardone RL, Padyana AK, Petersen MC, Judd ET, Hayes S, 
            Tong S, Barber KW, Apostolidi M, Abulizi A, Sheetz JB, Kshitiz, 
            Aerni HR, Gross S, Kung C, Samuel VT, Shulman GI, Kibbey RG, 
            Rinehart J
  TITLE     Distinct Hepatic PKA and CDK Signaling Pathways Control 
            Activity-Independent Pyruvate Kinase Phosphorylation and Hepatic 
            Glucose Production.
  JOURNAL   Cell Rep. 2019 Dec 10;29(11):3394-3404.e9. doi: 
            10.1016/j.celrep.2019.11.009.
  PUBMED    31825824
REFERENCE   2  (bases 1 to 7277)
  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"; doi: 
            "10.1016/j.celrep.2019.11"; journalName: "Cell Rep"; date: 
            "2019-12-10- 10"; volume: "29"; issue: "11"; pages: "3394-3404.e9"
COMMENT     Sequence Label: MBP-rPKL S113TAG
FEATURES             Location/Qualifiers
     source          1..7277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(246..1124)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1151..1435
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1358..1377
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1469..2569
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     2543..2566
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2618..2629
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4262..4279
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4327..4344)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4497..4583
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4675..4702
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4778..4796)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4792..4839
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4936..5294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5023..5042)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5233..5254
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5319..5423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5424..5492,5493..6284)
                     /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(5642..5661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6455..7043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6944..6963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7197..7214
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctatgat atcgattttt atttgatgcc tggcagttcc ctactctcgc atggggagac
       61 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg
      121 accaccgcgc tactgccgcc aggcatctag catgctgcat aatgtgcctg tcaaatggac
      181 gaagcaggga ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt
      241 accaattatg acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac
      301 tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca
      361 aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc
      421 gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg
      481 aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca
      541 aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc
      601 atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc
      661 agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc
      721 ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg
      781 gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc
      841 cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc
      901 gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc
      961 ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata
     1021 aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta
     1081 aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg
     1141 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
     1201 tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa
     1261 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
     1321 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
     1381 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg
     1441 ttttttgggc taacaggagg aattacatat gaaaatcgaa gaaggtaaac tggtaatctg
     1501 gattaacggc gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga
     1561 taccggaatt aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt
     1621 tgcggcaact ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta
     1681 cgctcaatct ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta
     1741 tccgtttacc tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt
     1801 tgaagcgtta tcgctgattt ataacaagga tctgctgccg aacccgccaa aaacctggga
     1861 agagatcccg gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa
     1921 cctgcaagaa ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa
     1981 gtatgaaaac ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc
     2041 gggtctgacc ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta
     2101 ctccatcgca gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg
     2161 ggcatggtcc aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt
     2221 caagggtcaa ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag
     2281 tccgaacaaa gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct
     2341 ggaagcggtt aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga
     2401 gttggcgaaa gatccacgta ttgccgccac tatggaaaac gcccagaaag gtgaaatcat
     2461 gccgaacatc ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc
     2521 cgccagcggt cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgagctc
     2581 gaacaacaac aacaataaca ataacaacaa cctcgggatc gagggaaggc atatggaagg
     2641 tccggcaggt tacctgcgcc gtgcgagcgt tgcgcagctg acccaagaac tgggcaccgc
     2701 attcttccag caacaacagc tgccggcggc tatggctgac acgttcctgg aacacctgtg
     2761 cctgctggat attgattccc agccagtggc ggctcgcagc acttccatta tcgctaccat
     2821 tggtccagct tcccgttctg tcgatcgcct gaaagaaatg atcaaagcag gcatgaacat
     2881 cgctcgtctg aacttttctc acggttctca cgaataccac gctgaaagca tcgcgaatat
     2941 ccgtgaagct accgaatcct tcgcgaccta gccgctgagc taccgtccgg tagcaatcgc
     3001 gctggacacc aaaggcccgg aaattcgtac cggcgtgctg cagggtggtc cggaatccga
     3061 agttgaaatt gtgaaaggtt ctcaggtgct ggtgaccgtt gacccgaaat tccaaacgcg
     3121 tggcgatgca aaaactgtgt gggtagacta ccacaacatt actcgtgtgg tagcggtagg
     3181 tggccgtatc tacatcgatg atggcctgat ctctctggtg gttcagaaaa tcggtccgga
     3241 aggtctggtt accgaagttg agcacggtgg tatcctgggt tctcgtaaag gcgttaacct
     3301 gcctaacacg gaagttgacc tgccgggcct gagcgaacag gatctgctgg atctgcgttt
     3361 cggcgttcaa cacaacgtgg atattatctt cgcatccttc gttcgtaaag cgtctgacgt
     3421 gctggctgta cgtgatgcgc tgggtccaga aggtcagaat atcaagatca tctctaaaat
     3481 tgaaaatcac gaaggcgtga aaaaatttga cgaaatcctg gaagtttccg atggcattat
     3541 ggtggcgcgc ggcgacctgg gcattgaaat tccggccgag aaagttttcc tggctcaaaa
     3601 gatgatgatc ggtcgctgta acctggctgg caagccggta gtatgcgcga cccagatgct
     3661 ggagagcatg atcactaaag cccgcccaac tcgcgccgaa accagcgatg ttgcaaacgc
     3721 tgtgctggac ggtgcagatt gcattatgct gtctggcgaa accgcaaaag gttcttttcc
     3781 ggttgaagct gttatgatgc aacacgcaat cgcgcgtgaa gctgaagcgg ccgtgtacca
     3841 ccgccagctg ttcgaagaac tgcgccgtgc tgcaccgctg tctcgcgacc cgactgaagt
     3901 taccgctatc ggcgcagttg aggcttcctt taaatgttgc gccgccgcta ttatcgtcct
     3961 gactaaaacc ggccgctccg cccaactgct gtctcagtat cgcccacgcg ctgctgttat
     4021 cgccgttacg cgcagcgcgc aggcagcccg tcaggttcac ctgtctcgtg gcgttttccc
     4081 gctgctgtac cgtgaaccgc ctgaagcgat ctgggccgac gatgtcgatc gtcgtgttca
     4141 gtttggtatt gaatccggca aactgcgtgg tttcctgcgt gttggtgacc tggttatcgt
     4201 cgttactggt tggcgtccgg gttctggtta taccaacatt atgcgcgttc tgtctgtgtc
     4261 ccaccaccat caccaccact aagcttgagc tcttggctgt tttggcggat gagagaagat
     4321 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
     4381 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
     4441 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     4501 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     4561 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     4621 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
     4681 ccatcctgac ggatggcctt tttgcgtttc tacaaactcc tcgagcttga tccggctgct
     4741 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     4801 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     4861 ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4921 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4981 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5041 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5101 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5161 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5221 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5281 attcttttga tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5401 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5461 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5521 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5581 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5641 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5701 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5761 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5821 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5881 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5941 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6001 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6061 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6121 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6181 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6241 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6601 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7081 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7141 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7201 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     7261 ggccgattca ttaatgc
//
