LOCUS       sequence-322793-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TNF and ST6GAL1 swapped chimera with the cytosolic tail 
            from TNF and TMD and luminal domain from ST6GAL1 in mammalian cells 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  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 5977)
  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: NSS-GFP
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1922..1931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1928..1930,1931..1933,1934..2647)
                     /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(1973..1994)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2769..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2806..2825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2860..2879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2897..3352)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3034..3055)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3246..3265
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3379..3483
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3485..3842
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3485..3676
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3508..3528)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3693..3828
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3877..4671
                     /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             3877..4671
                     /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(3931..3950)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4541..4560
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4859..4878)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4903..4950
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5279..5867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5768..5787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctaccggac tcagatctcg aggccaccat gtctaccgag tctatgatta gggacgtgga
      661 actggctgag gaggcactgc ccaaaaaaac cggcggacca cagggctcta ggagattcag
      721 ctgctgcgtc ctggtctttc ttctgtttgc agtcatctgt gtgtggaagg aaaagaagaa
      781 agggagttac tatgattcct ttaaattgca aaccaaggaa ttccaggtgt taaagagtct
      841 ggggaaattg gccatggggt ctgattccca gtctgtatcc tcaagcagca cccaggaccc
      901 ccacaggggc cgccagaccc tcggcagtct cagaggccta gccaaggcca aaccagaggc
      961 ctccttccag gtgtggaaca aggacagctc ttccaaaaac cttatcccta ggctgcaaaa
     1021 gatctggaag aattacctaa gcatgaacaa gtacaaagtg tcctacaagg ggccaggacc
     1081 aggcatcaag ttcagtgcag aggccctgcg ctgccacctc cgggaccatg tgaatgtatc
     1141 catggtagag gtcacagatt ttcccttcaa tacctctgaa tgggagggtt atctgcccaa
     1201 ggagagcatt aggaccaagg ctgggccttg gggcaggtgt gctgttgtgt cgtcagcggg
     1261 atctctgaag tcctcccaac taggcagaga aatcgatgat catgacgcag tcctgaggtt
     1321 taatggggca cccacagcca acttccaaca agatgtgggc acaaaaacta ccattcgcct
     1381 gatgaactct cagttggtta ccacagagaa gcgcttcctc aaagacagtt tgtacaatga
     1441 aggaatccta attgtatggg acccatctgt ataccactca gatatcccaa agtggtacca
     1501 gaatccggat tataatttct ttaacaacta caagacttat cgtaagctgc accccaatca
     1561 gcccttttac atcctcaagc cccagatgcc ttgggagcta tgggacattc ttcaagaaat
     1621 ctccccagaa gagattcagc caaacccccc atcctctggg atgcttggta tcatcatcat
     1681 gatgacgctg tgtgaccagg tggatattta tgagttcctc ccatccaagc gcaagactga
     1741 cgtgtgctac tactaccaga agttcttcga tagtgcctgc acgatgggtg cctaccaccc
     1801 gctgctctat gagaagaatt tggtgaagca tctcaaccag ggcacagatg aggacatcta
     1861 cctgcttgga aaagccacac tgcctggctt ccggaccatt cactgccggg atccaccggt
     1921 cgccaccatg gtgtccaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1981 gctggacggc gacgtaaacg gccacaagtt ctccgtgcgg ggcgagggcg agggcgatgc
     2041 caccaacggc aagctgaccc tgaagttcat cagcaccacc ggcaagctgc ccgtgccctg
     2101 gcccaccctc gtgaccaccc tgacctacgg cgtgcagagc ttctcccgct accccgacca
     2161 catgaagcgc cacgacttct tcaagagcgc catgcccgaa ggctacgtcc aggagcgcac
     2221 catctccttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga
     2281 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2341 ggggcacaag ctggagtaca acttcaactc ccacaacgtc tatatcaccg ccgacaagca
     2401 gaagaacggc atcaaggcca acttcaagat ccgccacaac gtggaggacg gctccgtgca
     2461 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2521 caaccactac ctgtccaccc agtccaagct gtccaaagac cccaacgaga agcgcgatca
     2581 catggtcctt ctggaattcg tgaccgccgc cgggatcact cacggcatgg acgagctgta
     2641 caagtaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     2701 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2761 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2881 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2941 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3001 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3061 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3121 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3181 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3241 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3301 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3361 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3421 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3481 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3541 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3601 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3661 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3721 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3781 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3841 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3901 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3961 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4021 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4081 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4141 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4201 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4261 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4321 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4381 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4441 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4501 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4561 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4621 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4681 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4741 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4801 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4861 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4921 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4981 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5041 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5101 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5161 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5221 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5281 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5341 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5401 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5461 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5521 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5581 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5641 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5701 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5761 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5821 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5881 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5941 tatcccctga ttctgtggat aaccgtatta ccgccat
//
