LOCUS       sequence-322799-        6007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TNF and ST6GAL1 swapped chimera with the cytosolic tail 
            and 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 6007)
  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 6007)
  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: NNS-GFP
FEATURES             Location/Qualifiers
     source          1..6007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        46..349
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        350..553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     504..524
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1933..1942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1939..1941,1942..1944,1945..2658)
                     /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(1984..2005)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2780..2901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2817..2836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2871..2890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2908..3363)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3045..3066)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3257..3276
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3390..3494
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3496..3853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3496..3687
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3519..3539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3704..3839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3888..4682
                     /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             3888..4682
                     /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(3942..3961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4552..4571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4870..4889)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4914..4961
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5290..5878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5779..5798
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
       61 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      121 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      181 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      241 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      301 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      361 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      421 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      481 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      541 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      601 gaggccacca tgtctaccga gtctatgatt agggacgtgg aactggctga ggaggcactg
      661 cccaaaaaaa ccggcggacc acagggctct aggagatgtc tgtttctgtc tctgttctct
      721 tttctcatcg tggctggcgc tacaacactc ttctgtctgc tccatttcgg cgtgattaag
      781 gaaaagaaga aagggagtta ctatgattcc tttaaattgc aaaccaagga attccaggtg
      841 ttaaagagtc tggggaaatt ggccatgggg tctgattccc agtctgtatc ctcaagcagc
      901 acccaggacc cccacagggg ccgccagacc ctcggcagtc tcagaggcct agccaaggcc
      961 aaaccagagg cctccttcca ggtgtggaac aaggacagct cttccaaaaa ccttatccct
     1021 aggctgcaaa agatctggaa gaattaccta agcatgaaca agtacaaagt gtcctacaag
     1081 gggccaggac caggcatcaa gttcagtgca gaggccctgc gctgccacct ccgggaccat
     1141 gtgaatgtat ccatggtaga ggtcacagat tttcccttca atacctctga atgggagggt
     1201 tatctgccca aggagagcat taggaccaag gctgggcctt ggggcaggtg tgctgttgtg
     1261 tcgtcagcgg gatctctgaa gtcctcccaa ctaggcagag aaatcgatga tcatgacgca
     1321 gtcctgaggt ttaatggggc acccacagcc aacttccaac aagatgtggg cacaaaaact
     1381 accattcgcc tgatgaactc tcagttggtt accacagaga agcgcttcct caaagacagt
     1441 ttgtacaatg aaggaatcct aattgtatgg gacccatctg tataccactc agatatccca
     1501 aagtggtacc agaatccgga ttataatttc tttaacaact acaagactta tcgtaagctg
     1561 caccccaatc agccctttta catcctcaag ccccagatgc cttgggagct atgggacatt
     1621 cttcaagaaa tctccccaga agagattcag ccaaaccccc catcctctgg gatgcttggt
     1681 atcatcatca tgatgacgct gtgtgaccag gtggatattt atgagttcct cccatccaag
     1741 cgcaagactg acgtgtgcta ctactaccag aagttcttcg atagtgcctg cacgatgggt
     1801 gcctaccacc cgctgctcta tgagaagaat ttggtgaagc atctcaacca gggcacagat
     1861 gaggacatct acctgcttgg aaaagccaca ctgcctggct tccggaccat tcactgccgg
     1921 gatccaccgg tcgccaccat ggtgtccaag ggcgaggagc tgttcaccgg ggtggtgccc
     1981 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tctccgtgcg gggcgagggc
     2041 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tcagcaccac cggcaagctg
     2101 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagag cttctcccgc
     2161 taccccgacc acatgaagcg ccacgacttc ttcaagagcg ccatgcccga aggctacgtc
     2221 caggagcgca ccatctcctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     2281 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2341 ggcaacatcc tggggcacaa gctggagtac aacttcaact cccacaacgt ctatatcacc
     2401 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa cgtggaggac
     2461 ggctccgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2521 ctgctgcccg acaaccacta cctgtccacc cagtccaagc tgtccaaaga ccccaacgag
     2581 aagcgcgatc acatggtcct tctggaattc gtgaccgccg ccgggatcac tcacggcatg
     2641 gacgagctgt acaagtaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     2701 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     2761 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     2821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2881 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     2941 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3001 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3061 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3121 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3181 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3241 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3301 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3361 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3481 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3541 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3601 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3721 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3781 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3841 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3901 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3961 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4021 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4081 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4141 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4201 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4261 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4321 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4381 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4441 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4501 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4561 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4621 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4681 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4741 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4801 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     4861 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     4921 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     4981 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5041 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5101 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5161 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5221 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5281 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5341 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5401 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5461 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5521 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5581 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5641 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5701 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5761 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5821 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5881 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5941 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     6001 aatagta
//
