LOCUS       sequence-322796-        5308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TNF and ST6GAL1 swapped chimera with the cytosolic tail 
            and 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 5308)
  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 5308)
  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: SSN-GFP
FEATURES             Location/Qualifiers
     source          1..5308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        48..351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        352..555
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     506..526
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1236..1245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1242..1244,1245..1247,1248..1961)
                     /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(1287..1308)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2083..2204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2120..2139)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2174..2193
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2211..2666)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2348..2369)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2560..2579
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2693..2797
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2799..3156
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2799..2990
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2822..2842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3007..3142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3191..3985
                     /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             3191..3985
                     /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(3245..3264)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3855..3874
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4173..4192)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4217..4264
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4593..5181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5082..5101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
       61 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      121 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      181 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      241 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      301 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      361 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      421 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      481 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      541 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc
      601 tcgaggccac catgattcac accaacctga agaaaaagtt cagctgctgc gtcctggtct
      661 ttcttctgtt tgcagtcatc tgtgtgtggg gaccacagcg agaggaattt ccccgggatc
      721 tgtcactcat ttcaccactg gcacaggctg tccgatcttc atctcggact ccatccgaca
      781 aacctgtcgc ccatgtcgtc gccaacccac aggccgaggg ccagctccag tggctcaata
      841 ggagggcaaa cgctctgctc gccaatggcg tggaactccg ggataaccag ctcgtcgtgc
      901 ctagtgaggg actgtacctc atctactccc aggtgctgtt taagggccag ggatgtcctt
      961 ctacacatgt gctgctcaca cacacaattt cacggatcgc cgtgtcttac cagactaaag
     1021 tcaatctgct ctctgccatc aaatccccat gtcagcggga aacacctgag ggcgctgagg
     1081 ctaaaccttg gtacgaaccc atctacctcg gaggcgtgtt ccagctggag aagggcgata
     1141 gactgagtgc tgagatcaat cgacccgact acctcgactt tgccgaatct ggccaggtct
     1201 actttggcat cattgctctg cgggatccac cggtcgccac catggtgtcc aagggcgagg
     1261 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1321 agttctccgt gcggggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt
     1381 tcatcagcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1441 acggcgtgca gagcttctcc cgctaccccg accacatgaa gcgccacgac ttcttcaaga
     1501 gcgccatgcc cgaaggctac gtccaggagc gcaccatctc cttcaaggac gacggcacct
     1561 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1621 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca
     1681 actcccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1741 agatccgcca caacgtggag gacggctccg tgcagctcgc cgaccactac cagcagaaca
     1801 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgtcc acccagtcca
     1861 agctgtccaa agaccccaac gagaagcgcg atcacatggt ccttctggaa ttcgtgaccg
     1921 ccgccgggat cactcacggc atggacgagc tgtacaagta agcggccgcg actctagatc
     1981 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2041 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2101 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2161 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     2221 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2281 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2341 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2401 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2461 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2521 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2581 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2641 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     2701 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2761 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     2821 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2881 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3001 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3061 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3121 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3181 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3241 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3301 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3361 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3421 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3481 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3541 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3601 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3661 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     3721 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3781 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3841 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3901 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3961 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4021 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4081 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4141 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4201 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4261 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4321 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4381 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4441 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4501 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5281 attaccgcca tgcattagtt attaatag
//
