LOCUS       sequence-392656-        6444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Eucaryotic expression plasmid for chimeric cytokine of IL-11 and LIF
            (GIL-11) with a TwinStrep-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6444)
  AUTHORS   Rafii P, Seibel C, Weitz HT, Ettich J, Minafra AR, Petzsch P, Lang 
            A, Floss DM, Behnke K, Kohrer K, Moll JM, Scheller J
  TITLE     Cytokimera GIL-11 rescued IL-6R deficient mice from partial 
            hepatectomy-induced death by signaling via non-natural 
            gp130:LIFR:IL-11R complexes.
  JOURNAL   Commun Biol. 2023 Apr 15;6(1):418. doi: 10.1038/s42003-023-04768-4.
  PUBMED    37061565
REFERENCE   2  (bases 1 to 6444)
  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: "Commun 
            Biol."; date: "2023-04-15"; volume: "6(1):418. doi"; pages: " 
            10.1038/s42003-023-04768-4"
COMMENT     Sequence Label: pcDNA3.1 GIL-11 myc TEV TwinStrep
FEATURES             Location/Qualifiers
     source          1..6444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     866..885
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        866..884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             992..1021
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1613..1633
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             join(1643..1648,1649..1672,1673..1708,1709..1732)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     primer_bind     complement(2038..2055)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2044..2268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2314..2742
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2401..2420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2611..2632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2751..2771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2756..3085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2936..3071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2998..3017
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3152..3946
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3206..3225)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3816..3835
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4120..4241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4157..4176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4211..4230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4290..4306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4290..4306)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4303..4325)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4314..4330
                     /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(4338..4344,4345..4362,4363..4368))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4383..4404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4521..4538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4692..5280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4772..4791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5451..6242,6243..6311))
                     /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     6074..6093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6312..6416)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      841 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      901 agcgtttaaa cttaagacca tgagcagcag ctgctcaggg ctgagcaggg tcctggtggc
      961 cgtggctaca gccctggtgt ctgcctcctc cgaacaaaaa ctcatctcag aagaggatct
     1021 gaagcttgga ccaccccctg gacctcccag ggtgagccca gaccctagag ccgagctgga
     1081 cagcacagtg ctgctgacca ggtccctgct ggccgataca agacagctgg ccatcctgta
     1141 ctacacagcc cagggcgagc ccttccctaa taacctggat aagctgtgtg gccctaatgt
     1201 gacagacagc ctgcctacac tggccatgag cgccggcgcc ctgggagctc tgcagctgcc
     1261 tggagtgctg acaagactga gggccgacct gctgagctac ctgagacacg tgcagtggat
     1321 caccagagat cagaagatcc tgaatcctag cgccctgagc ctgcactcca agctgaatac
     1381 cctgcaggcc aggctggaca gactgctgag gaggctgcag ctgctgatga gcagactggc
     1441 cctgccccag cctcccccag accctcctgc tccacccctg gctacatccg gcaaggacgt
     1501 gttccagaag aagaagctgg gctgccagct gctgggcggc ctgcatctga cactggattg
     1561 ggccgtgaga ggcctgctgc tgctgaagac cagactgggc ggcggcggct ccgagaacct
     1621 gtacttccag agcgcggccg caagcgcgtg gagccatccg cagtttgaaa aaggcggcgg
     1681 cagcggcggc ggcagcggcg gcagcgcgtg gagccatccg cagtttgaaa aataagttta
     1741 aaccgaattc tgcagatatc gctgtggagt gggagtctaa tggccagccc gagaacaatt
     1801 acaagaccac ccctcctgtg ctggattctg acggctcctt cttcctgtac tccaaactga
     1861 ccgtggacaa gtctagatgg cagcagggca acgtgttctc ttgttccgtg atgcacgagg
     1921 ctctgcacaa tcactatacc cagaagtccc tgtctctgtc tcatggcaag tgaggatcca
     1981 ctagtccagt gtggtggaat ttcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     2041 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2101 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     2161 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     2221 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     2281 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2341 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2401 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2461 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2521 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2581 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2641 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2701 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2761 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2821 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2881 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2941 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3001 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3061 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3121 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3181 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3241 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3301 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3361 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3421 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3481 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3541 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3601 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3661 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3721 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3781 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3841 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3901 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3961 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4021 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4081 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4261 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4321 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4381 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4441 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4501 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4561 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4621 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4681 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4741 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4801 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4861 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4921 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4981 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5041 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5101 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5161 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5221 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5281 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5341 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5401 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5461 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5521 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5581 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5641 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5701 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5761 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5821 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5881 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5941 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6001 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6061 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6121 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6181 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6241 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6301 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6361 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6421 tttccccgaa aagtgccacc tgac
//
