LOCUS       sequence-322795-        5947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TNF and ST6GAL1 swapped chimera with the cytosolic tail 
            from ST6GAL1, TMD from TNF and luminal domain from ST6GAL1 in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5947)
  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 5947)
  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: SNS-GFP
FEATURES             Location/Qualifiers
     source          1..5947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1890..1899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1896..1898,1899..1901,1902..2615)
                     /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(1941..1962)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2737..2858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2774..2793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2828..2847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2865..3320)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3002..3023)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3214..3233
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3347..3451
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3453..3810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3453..3644
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3476..3496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3661..3796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3845..4639
                     /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             3845..4639
                     /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(3899..3918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4509..4528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4827..4846)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4871..4918
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5247..5835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5736..5755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      601 taccggactc agatctcgag gccaccatga ttcacaccaa cctgaagaaa aagtgtctgt
      661 ttctgtctct gttctctttt ctcatcgtgg ctggcgctac aacactcttc tgtctgctcc
      721 atttcggcgt gattaaggaa aagaagaaag ggagttacta tgattccttt aaattgcaaa
      781 ccaaggaatt ccaggtgtta aagagtctgg ggaaattggc catggggtct gattcccagt
      841 ctgtatcctc aagcagcacc caggaccccc acaggggccg ccagaccctc ggcagtctca
      901 gaggcctagc caaggccaaa ccagaggcct ccttccaggt gtggaacaag gacagctctt
      961 ccaaaaacct tatccctagg ctgcaaaaga tctggaagaa ttacctaagc atgaacaagt
     1021 acaaagtgtc ctacaagggg ccaggaccag gcatcaagtt cagtgcagag gccctgcgct
     1081 gccacctccg ggaccatgtg aatgtatcca tggtagaggt cacagatttt cccttcaata
     1141 cctctgaatg ggagggttat ctgcccaagg agagcattag gaccaaggct gggccttggg
     1201 gcaggtgtgc tgttgtgtcg tcagcgggat ctctgaagtc ctcccaacta ggcagagaaa
     1261 tcgatgatca tgacgcagtc ctgaggttta atggggcacc cacagccaac ttccaacaag
     1321 atgtgggcac aaaaactacc attcgcctga tgaactctca gttggttacc acagagaagc
     1381 gcttcctcaa agacagtttg tacaatgaag gaatcctaat tgtatgggac ccatctgtat
     1441 accactcaga tatcccaaag tggtaccaga atccggatta taatttcttt aacaactaca
     1501 agacttatcg taagctgcac cccaatcagc ccttttacat cctcaagccc cagatgcctt
     1561 gggagctatg ggacattctt caagaaatct ccccagaaga gattcagcca aaccccccat
     1621 cctctgggat gcttggtatc atcatcatga tgacgctgtg tgaccaggtg gatatttatg
     1681 agttcctccc atccaagcgc aagactgacg tgtgctacta ctaccagaag ttcttcgata
     1741 gtgcctgcac gatgggtgcc taccacccgc tgctctatga gaagaatttg gtgaagcatc
     1801 tcaaccaggg cacagatgag gacatctacc tgcttggaaa agccacactg cctggcttcc
     1861 ggaccattca ctgccgggat ccaccggtcg ccaccatggt gtccaagggc gaggagctgt
     1921 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttct
     1981 ccgtgcgggg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatca
     2041 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2101 tgcagagctt ctcccgctac cccgaccaca tgaagcgcca cgacttcttc aagagcgcca
     2161 tgcccgaagg ctacgtccag gagcgcacca tctccttcaa ggacgacggc acctacaaga
     2221 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2281 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ttcaactccc
     2341 acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc
     2401 gccacaacgt ggaggacggc tccgtgcagc tcgccgacca ctaccagcag aacaccccca
     2461 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gtccacccag tccaagctgt
     2521 ccaaagaccc caacgagaag cgcgatcaca tggtccttct ggaattcgtg accgccgccg
     2581 ggatcactca cggcatggac gagctgtaca agtaagcggc cgcgactcta gatcataatc
     2641 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2701 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2761 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2821 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2881 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2941 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3001 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3061 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3121 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3181 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3241 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3301 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3361 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3421 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3541 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3721 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3781 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3841 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3901 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3961 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4021 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4081 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4141 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4201 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4261 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4321 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4381 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4441 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4501 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4561 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4621 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4681 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4741 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4801 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4861 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4921 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4981 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5041 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5101 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5161 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5221 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5281 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5341 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5401 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5461 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5521 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5581 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5641 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5701 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5761 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5821 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5881 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5941 gccatgc
//
