LOCUS       sequence-322789-        6958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with the cytosolic tail 
            from TFR, TMD from ST6GAL1, and luminal domain from TFR in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6958)
  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 6958)
  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: FSF-GFP
FEATURES             Location/Qualifiers
     source          1..6958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(1562..1573)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2918..2927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2924..2926,2927..2929,2930..3643)
                     /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(2969..2990)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3765..3886
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3802..3821)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3856..3875
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3893..4348)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4030..4051)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4242..4261
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4375..4479
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4481..4838
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4481..4672
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4504..4524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4689..4824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4873..5667
                     /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             4873..5667
                     /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(4927..4946)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5537..5556
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5855..5874)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5899..5946
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6275..6863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6764..6783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      601 gtcagatccg ctagcgctac cggactcaga tctcgaggcc accatgatgg atcaagctag
      661 atcagcattc tctaacttgt ttggtggaga accattgtca tatacccggt tcagcctggc
      721 tcggcaagta gatggcgata acagtcatgt ggagatgaaa cttgctgtag atgaagaaga
      781 aaatgctgac aataacacaa aggccaatgt cacaaaacca aaaaggttca gctgctgcgt
      841 cctggtcttt cttctgtttg cagtcatctg tgtgtggaag gaaaagtgta aaggggtaga
      901 accaaaaact gagtgtgaga gactggcagg aaccgagtct ccagtgaggg aggagccagg
      961 agaggacttc cctgcagcac gtcgcttata ttgggatgac ctgaagagaa agttgtcgga
     1021 gaaactggac agcacagact tcaccagcac catcaagctg ctgaatgaaa attcatatgt
     1081 ccctcgtgag gctggatctc aaaaagatga aaatcttgcg ttgtatgttg aaaatcaatt
     1141 tcgtgaattt aaactcagca aagtctggcg tgatcaacat tttgttaaga ttcaggtcaa
     1201 agacagcgct caaaactcgg tgatcatagt tgataagaac ggtagacttg tttacctggt
     1261 ggagaatcct gggggttatg tggcgtatag taaggctgca acagttactg gtaaactggt
     1321 ccatgctaat tttggtacta aaaaagattt tgaggattta tacactcctg tgaatggatc
     1381 tatagtgatt gtcagagcag ggaaaatcac ctttgcagaa aaggttgcaa atgctgaaag
     1441 cttaaatgca attggtgtgt tgatatacat ggaccagact aaatttccca ttgttaacgc
     1501 agaactttca ttctttggac atgctcatct ggggacaggt gacccttaca cacctggatt
     1561 cccttccttc aatcacactc agtttccacc atctcggtca tcaggattgc ctaatatacc
     1621 tgtccagaca atctccagag ctgctgcaga aaagctgttt gggaatatgg aaggagactg
     1681 tccctctgac tggaaaacag actctacatg taggatggta acctcagaaa gcaagaatgt
     1741 gaagctcact gtgagcaatg tgctgaaaga gataaaaatt cttaacatct ttggagttat
     1801 taaaggcttt gtagaaccag atcactatgt tgtagttggg gcccagagag atgcatgggg
     1861 ccctggagct gcaaaatccg gtgtaggcac agctctccta ttgaaacttg cccagatgtt
     1921 ctcagatatg gtcttaaaag atgggtttca gcccagcaga agcattatct ttgccagttg
     1981 gagtgctgga gactttggat cggttggtgc cactgaatgg ctagagggat acctttcgtc
     2041 cctgcattta aaggctttca cttatattaa tctggataaa gcggttcttg gtaccagcaa
     2101 cttcaaggtt tctgccagcc cactgttgta tacgcttatt gagaaaacaa tgcaaaatgt
     2161 gaagcatccg gttactgggc aatttctata tcaggacagc aactgggcca gcaaagttga
     2221 gaaactcact ttagacaatg ctgctttccc tttccttgca tattctggaa tcccagcagt
     2281 ttctttctgt ttttgcgagg acacagatta tccttatttg ggtaccacca tggacaccta
     2341 taaggaactg attgagagga ttcctgagtt gaacaaagtg gcacgagcag ctgcagaggt
     2401 cgctggtcag ttcgtgatta aactaaccca tgatgttgaa ttgaacctgg actatgagag
     2461 gtacaacagc caactgcttt catttgtgag ggatctgaac caatacagag cagacataaa
     2521 ggaaatgggc ctgagtttac agtggctgta ttctgctcgt ggagacttct tccgtgctac
     2581 ttccagacta acaacagatt tcgggaatgc tgagaaaaca gacagatttg tcatgaagaa
     2641 actcaatgat cgtgtcatga gagtggagta tcacttcctc tctccctacg tatctccaaa
     2701 agagtctcct ttccgacatg tcttctgggg ctccggctct cacacgctgc cagctttact
     2761 ggagaacttg aaactgcgta aacaaaataa cggtgctttt aatgaaacgc tgttcagaaa
     2821 ccagttggct ctagctactt ggactattca gggagctgca aatgccctct ctggtgacgt
     2881 ttgggacatt gacaatgagt ttcgggatcc accggtcgcc accatggtgt ccaagggcga
     2941 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     3001 caagttctcc gtgcggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     3061 gttcatcagc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3121 ctacggcgtg cagagcttct cccgctaccc cgaccacatg aagcgccacg acttcttcaa
     3181 gagcgccatg cccgaaggct acgtccagga gcgcaccatc tccttcaagg acgacggcac
     3241 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3301 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     3361 caactcccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     3421 caagatccgc cacaacgtgg aggacggctc cgtgcagctc gccgaccact accagcagaa
     3481 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctgt ccacccagtc
     3541 caagctgtcc aaagacccca acgagaagcg cgatcacatg gtccttctgg aattcgtgac
     3601 cgccgccggg atcactcacg gcatggacga gctgtacaag taagcggccg cgactctaga
     3661 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3721 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3781 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3841 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3901 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3961 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4021 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4081 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4141 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4201 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4261 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4321 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4381 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4441 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4501 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4561 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4621 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4681 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4741 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4801 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4861 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4921 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4981 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5041 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5101 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5161 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5221 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5281 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5341 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5401 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5461 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5521 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5581 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5641 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5701 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5761 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5821 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5881 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5941 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6001 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6061 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6121 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6181 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6241 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6301 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6361 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6421 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6481 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6541 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6601 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6661 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6721 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6781 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6841 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6901 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataa
//
