LOCUS       sequence-213785-        5327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human interleukin-6 (IL6) fused to GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5327)
  AUTHORS   Verboogen DRJ, Revelo NH, Ter Beest M, van den Bogaart G
  TITLE     Interleukin-6 secretion is limited by self-signaling in endosomes.
  JOURNAL   J Mol Cell Biol. 2018 Jul 16. pii: 5054600. doi: 
            10.1093/jmcb/mjy038.
  PUBMED    30016456
REFERENCE   2  (bases 1 to 5327)
  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: "J Mol Cell 
            Biol. 2018 Jul 16. pii: 5054600. doi: 10.1093/jmcb/mjy038."
COMMENT     Sequence Label: pEGFP-N1-IL6
FEATURES             Location/Qualifiers
     source          1..5327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        783..1086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1087..1290
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1241..1261
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1989..1998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1995..1997,1998..2000,2001..2714)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1995..1997,1998..2000,2001..2711)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2040..2061)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2301..2320)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2648..2669
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2837..2958
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2874..2893)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2928..2947
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2965..3420)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3102..3123)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3314..3333
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3447..3551
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3553..3910
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3553..3744
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3576..3596)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3761..3896
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3945..4739
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3945..4739
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3999..4018)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4609..4628
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4927..4946)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4971..5018
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
      721 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      781 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      841 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      901 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      961 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1021 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1081 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     1141 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     1201 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     1261 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
     1321 taccggactc agatctcgag atgaacagct tcagcaccag cgccttcggc cccgtggcct
     1381 tcagcctggg cctgctgctg gtgctgcccg ccgccttccc cgcccccgtg ccccccggcg
     1441 aggacagcaa ggacgtggcc gccccccacc gccagcccct gaccagcagc gagcgcatcg
     1501 acaagcagat ccgctacatc ctggacggca tcagcgccct gcgcaaggag acctgcaaca
     1561 agagcaacat gtgcgagagc agcaaggagg ccctggccga gaacaacctg aacctgccca
     1621 agatggccga gaaggacggc tgcttccaga gcggcttcaa cgaggagacc tgcctggtga
     1681 agatcatcac cggcctgctg gagttcgagg tgtacctgga gtacctgcag aaccgcttcg
     1741 agagcagcga ggagcaggcc cgcgccgtgc agatgagcac caaggtgctg atccagttcc
     1801 tgcagaagaa ggccaagaac ctggacgcca tcaccacccc cgaccccacc accaacgcca
     1861 gcctgctgac caagctgcag gcccagaacc agtggctgca ggacatgacc acccacctga
     1921 tcctgcgcag cttcaaggag ttcctgcaga gcagcctgcg cgccctgcgc cagatggatc
     1981 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2041 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2101 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2161 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2221 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2281 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2341 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2401 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2461 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2521 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2581 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2641 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2701 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2761 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2821 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2881 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2941 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3001 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3061 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3121 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3181 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3241 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3301 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3361 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3421 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3481 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3541 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3601 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3661 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3721 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3781 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3841 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3901 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3961 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4021 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4081 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4141 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4201 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4261 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4321 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4381 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4441 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4501 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4561 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4621 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4681 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4741 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4801 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4861 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4921 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4981 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5041 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5101 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5161 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtag
//
