LOCUS       sequence-322798-        5368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TNF and ST6GAL1 swapped chimera with the cytosolic tail 
            from TNF, TMD from ST6GAL1 and luminal domain from TNF in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5368)
  AUTHORS   Sun X, Mahajan D, Chen B, Song Z, Lu L
  TITLE     A quantitative study of the Golgi retention of glycosyltransferases.
  JOURNAL   J Cell Sci. 2021 Oct 15;134(20). pii: 272560. doi: 
            10.1242/jcs.258564. Epub 2021 Oct 21.
  PUBMED    34533190
REFERENCE   2  (bases 1 to 5368)
  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 Cell 
            Sci."; date: "2021-10-15"; volume: "134(20). pii: 272560. doi"; 
            pages: " 10.1242/jcs.258564. Epub 2021 Oct 21"
COMMENT     Sequence Label: NSN-GFP
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1316..1325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1322..1324,1325..1327,1328..2041)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1367..1388)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2163..2284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2200..2219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2254..2273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2291..2746)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2428..2449)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2640..2659
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2773..2877
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2879..3236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2879..3070
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2902..2922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3087..3222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3271..4065
                     /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             3271..4065
                     /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(3325..3344)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3935..3954
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4253..4272)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4297..4344
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4673..5261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5162..5181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgaggccac catgtctacc gagtctatga ttagggacgt
      661 ggaactggct gaggaggcac tgcccaaaaa aaccggcgga ccacagggct ctaggagatt
      721 cagctgctgc gtcctggtct ttcttctgtt tgcagtcatc tgtgtgtggg gaccacagcg
      781 agaggaattt ccccgggatc tgtcactcat ttcaccactg gcacaggctg tccgatcttc
      841 atctcggact ccatccgaca aacctgtcgc ccatgtcgtc gccaacccac aggccgaggg
      901 ccagctccag tggctcaata ggagggcaaa cgctctgctc gccaatggcg tggaactccg
      961 ggataaccag ctcgtcgtgc ctagtgaggg actgtacctc atctactccc aggtgctgtt
     1021 taagggccag ggatgtcctt ctacacatgt gctgctcaca cacacaattt cacggatcgc
     1081 cgtgtcttac cagactaaag tcaatctgct ctctgccatc aaatccccat gtcagcggga
     1141 aacacctgag ggcgctgagg ctaaaccttg gtacgaaccc atctacctcg gaggcgtgtt
     1201 ccagctggag aagggcgata gactgagtgc tgagatcaat cgacccgact acctcgactt
     1261 tgccgaatct ggccaggtct actttggcat cattgctctg cgggatccac cggtcgccac
     1321 catggtgtcc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1381 cggcgacgta aacggccaca agttctccgt gcggggcgag ggcgagggcg atgccaccaa
     1441 cggcaagctg accctgaagt tcatcagcac caccggcaag ctgcccgtgc cctggcccac
     1501 cctcgtgacc accctgacct acggcgtgca gagcttctcc cgctaccccg accacatgaa
     1561 gcgccacgac ttcttcaaga gcgccatgcc cgaaggctac gtccaggagc gcaccatctc
     1621 cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1681 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1741 caagctggag tacaacttca actcccacaa cgtctatatc accgccgaca agcagaagaa
     1801 cggcatcaag gccaacttca agatccgcca caacgtggag gacggctccg tgcagctcgc
     1861 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1921 ctacctgtcc acccagtcca agctgtccaa agaccccaac gagaagcgcg atcacatggt
     1981 ccttctggaa ttcgtgaccg ccgccgggat cactcacggc atggacgagc tgtacaagta
     2041 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2101 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2161 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2221 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2281 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2341 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2401 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2461 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2521 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2581 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2641 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2701 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2761 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2821 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2881 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2941 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3001 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3061 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3121 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3181 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3241 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3421 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3781 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4201 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4261 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4321 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4381 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4441 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4501 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4561 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4621 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4801 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5221 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5281 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5341 ctgattctgt ggataaccgt attaccgc
//
