LOCUS       sequence-234226-        4962 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 4962)
  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 4962)
  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..4962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1756..1765
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2200..2223
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2319..2338)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2384..2439
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2438..2457)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2798..2814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2798..2814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2811..2833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2822..2838
                     /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(2846..2852,2853..2870,2871..2876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2891..2912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3041..3058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3212..3800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3292..3311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3971..4762,4763..4831))
                     /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     4594..4613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4832..4936)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gcgcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttgttgt gctgtctcat cattttggca aagaattgcg gccgtctcag
     1741 gccgagttcg gtaccgccac catgggcatc cttcccagcc ctgggatgcc tgcgctgctc
     1801 tccctcgtga gccttctctc cgtgctgctg atgggttgcg tagctgaagg atccgcggac
     1861 agcccctcag cccctgtgaa cgtgaccgtc cggcacctca aggccaactc tgccgtggtc
     1921 agctgggatg tcctggagga tgaagtggtc attggctttg ccatctctca gcagaagaag
     1981 gatgtgcgga tgctccggtt cattcaggag gtgaacacca ccacccggtc ctgcgctctc
     2041 tgggacctgg aggaggacac agaatatatc gtccatgtgc aggccatctc catccaggga
     2101 cagagcccag ccagtgagcc tgtgctcttc aagaccccac gcgaggctga aaagatggcc
     2161 tcaaagaaca aagatgaggt gaccatgaag gaggaattcc atcaccacca tcatcaccat
     2221 cattgactcg agcgataatt cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     2281 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     2341 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     2401 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     2461 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgccata
     2521 tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga aacagccccc
     2581 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     2641 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     2701 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     2761 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2821 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2881 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2941 agtcgggaaa cctgtcgtgc cagcggatcg atccgctgca ttaatgaatc ggccaacgcg
     3001 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3061 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3121 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3181 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3241 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3301 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3361 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     3421 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3481 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3541 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3601 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3661 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3721 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4621 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4801 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4861 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4921 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tg
//
