LOCUS       sequence-239333-        4969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses in mammalian cells (HEK293):glycosylated and secreted 
            irisin-his.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  AUTHORS   Albrecht E, Norheim F, Thiede B, Holen T, Ohashi T, Schering L, Lee 
            S, Brenmoehl J, Thomas S, Drevon CA, Erickson HP, Maak S
  TITLE     Irisin - a myth rather than an exercise-inducible myokine.
  JOURNAL   Sci Rep. 2015 Mar 9;5:8889. doi: 10.1038/srep08889.
  PUBMED    25749243
REFERENCE   2  (bases 1 to 4969)
  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: "Sci Rep."; 
            date: "2015-03-9"; volume: "5:8889. doi"; pages: " 
            10.1038/srep08889"
COMMENT     Sequence Label: pHLSec2-irisin-his
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1761..1770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2205..2228
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2324..2343)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2389..2444
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2443..2462)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2805..2821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2805..2821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2818..2840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2829..2845
                     /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(2853..2859,2860..2877,2878..2883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2898..2919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3048..3065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3219..3807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3299..3318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3978..4769,4770..4838))
                     /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     4601..4620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4839..4943)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgcg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs grggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg grgggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagcgcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa ttgcggccgt
     1741 ctcaggccga gttcggtacc gccaccatgg gcatccttcc cagccctggg atgcctgcgc
     1801 tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct gaaggatccg
     1861 cggacagccc ctcagcccct gtgaacgtga ccgtccggca cctcaaggcc aactctgccg
     1921 tggtcagctg ggatgtcctg gaggatgaag tggtcattgg ctttgccatc tctcagcaga
     1981 agaaggatgt gcggatgctc cggttcattc aggaggtgaa caccaccacc cggtcctgcg
     2041 ctctctggga cctggaggag gacacagaat atatcgtcca tgtgcaggcc atctccatcc
     2101 agggacagag cccagccagt gagcctgtgc tcttcaagac cccacgcgag gctgaaaaga
     2161 tggcctcaaa gaacaaagat gaggtgacca tgaaggagga attccatcac caccatcatc
     2221 accatcattg actcgagcga taattcactc ctcaggtgca ggctgcctat cagaaggtgg
     2281 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     2341 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     2401 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     2461 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     2521 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     2581 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     2641 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     2701 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     2761 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     2821 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2881 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2941 gctttccagt cgggaaacct gtcgtgccag cggatcgatc cgctgcatta atgaatcggc
     3001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3901 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3961 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4021 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4081 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4141 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4201 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4261 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4321 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4381 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4441 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4501 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4561 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4621 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4681 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4741 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4801 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
