LOCUS       sequence-322781-        5947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with the cytosolic tail 
            from ST6GAL1, TMD from TFR 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: SFS-GFP
FEATURES             Location/Qualifiers
     source          1..5947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1887..1896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1893..1895,1896..1898,1899..2612)
                     /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(1938..1959)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2734..2855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2771..2790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2825..2844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2862..3317)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2999..3020)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3211..3230
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3344..3448
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3450..3807
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3450..3641
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3473..3493)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3658..3793
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3842..4636
                     /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             3842..4636
                     /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(3896..3915)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4506..4525
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4824..4843)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4868..4915
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5244..5832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5733..5752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggactcaga tctcgaggcc accatgattc acaccaacct gaagaaaaag tgtagtggaa
      661 gtatctgcta tgggactatt gctgtgatcg tctttttctt gattggattt atgattggct
      721 acttgggcta taaggaaaag aagaaaggga gttactatga ttcctttaaa ttgcaaacca
      781 aggaattcca ggtgttaaag agtctgggga aattggccat ggggtctgat tcccagtctg
      841 tatcctcaag cagcacccag gacccccaca ggggccgcca gaccctcggc agtctcagag
      901 gcctagccaa ggccaaacca gaggcctcct tccaggtgtg gaacaaggac agctcttcca
      961 aaaaccttat ccctaggctg caaaagatct ggaagaatta cctaagcatg aacaagtaca
     1021 aagtgtccta caaggggcca ggaccaggca tcaagttcag tgcagaggcc ctgcgctgcc
     1081 acctccggga ccatgtgaat gtatccatgg tagaggtcac agattttccc ttcaatacct
     1141 ctgaatggga gggttatctg cccaaggaga gcattaggac caaggctggg ccttggggca
     1201 ggtgtgctgt tgtgtcgtca gcgggatctc tgaagtcctc ccaactaggc agagaaatcg
     1261 atgatcatga cgcagtcctg aggtttaatg gggcacccac agccaacttc caacaagatg
     1321 tgggcacaaa aactaccatt cgcctgatga actctcagtt ggttaccaca gagaagcgct
     1381 tcctcaaaga cagtttgtac aatgaaggaa tcctaattgt atgggaccca tctgtatacc
     1441 actcagatat cccaaagtgg taccagaatc cggattataa tttctttaac aactacaaga
     1501 cttatcgtaa gctgcacccc aatcagccct tttacatcct caagccccag atgccttggg
     1561 agctatggga cattcttcaa gaaatctccc cagaagagat tcagccaaac cccccatcct
     1621 ctgggatgct tggtatcatc atcatgatga cgctgtgtga ccaggtggat atttatgagt
     1681 tcctcccatc caagcgcaag actgacgtgt gctactacta ccagaagttc ttcgatagtg
     1741 cctgcacgat gggtgcctac cacccgctgc tctatgagaa gaatttggtg aagcatctca
     1801 accagggcac agatgaggac atctacctgc ttggaaaagc cacactgcct ggcttccgga
     1861 ccattcactg ccgggatcca ccggtcgcca ccatggtgtc caagggcgag gagctgttca
     1921 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttctccg
     1981 tgcggggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatcagca
     2041 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     2101 agagcttctc ccgctacccc gaccacatga agcgccacga cttcttcaag agcgccatgc
     2161 ccgaaggcta cgtccaggag cgcaccatct ccttcaagga cgacggcacc tacaagaccc
     2221 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2281 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc aactcccaca
     2341 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     2401 acaacgtgga ggacggctcc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2461 gcgacggccc cgtgctgctg cccgacaacc actacctgtc cacccagtcc aagctgtcca
     2521 aagaccccaa cgagaagcgc gatcacatgg tccttctgga attcgtgacc gccgccggga
     2581 tcactcacgg catggacgag ctgtacaagt aagcggccgc gactctagat cataatcagc
     2641 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2701 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2761 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2821 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2881 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2941 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3001 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3061 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3121 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3181 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3241 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3301 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3361 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3421 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3481 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3541 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3601 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3661 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3721 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3781 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3841 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3901 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3961 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4021 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4081 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4141 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4201 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4261 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4321 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4381 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4441 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4501 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4561 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4621 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4681 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4741 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4801 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4861 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4921 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4981 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5041 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5101 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5161 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5221 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5281 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5341 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5401 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5461 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5521 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5581 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5641 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5701 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5761 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5821 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5881 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5941 atgcatt
//
