LOCUS       sequence-322817-        6802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with a truncated 
            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 6802)
  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 6802)
  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: F9SF-GFP
FEATURES             Location/Qualifiers
     source          1..6802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      625..634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1393..1404)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2749..2758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2755..2757,2758..2760,2761..3474)
                     /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(2800..2821)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3596..3717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3633..3652)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3687..3706
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3724..4179)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3861..3882)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4073..4092
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4206..4310
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4312..4669
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4312..4503
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4335..4355)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4520..4655
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4704..5498
                     /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             4704..5498
                     /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(4758..4777)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5368..5387
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5686..5705)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5730..5777
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6106..6694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6595..6614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgctaccgg actcagatct cgaggccacc atggccaatg tcacaaaacc aaaaaggttc
      661 agctgctgcg tcctggtctt tcttctgttt gcagtcatct gtgtgtggaa ggaaaagtgt
      721 aaaggggtag aaccaaaaac tgagtgtgag agactggcag gaaccgagtc tccagtgagg
      781 gaggagccag gagaggactt ccctgcagca cgtcgcttat attgggatga cctgaagaga
      841 aagttgtcgg agaaactgga cagcacagac ttcaccagca ccatcaagct gctgaatgaa
      901 aattcatatg tccctcgtga ggctggatct caaaaagatg aaaatcttgc gttgtatgtt
      961 gaaaatcaat ttcgtgaatt taaactcagc aaagtctggc gtgatcaaca ttttgttaag
     1021 attcaggtca aagacagcgc tcaaaactcg gtgatcatag ttgataagaa cggtagactt
     1081 gtttacctgg tggagaatcc tgggggttat gtggcgtata gtaaggctgc aacagttact
     1141 ggtaaactgg tccatgctaa ttttggtact aaaaaagatt ttgaggattt atacactcct
     1201 gtgaatggat ctatagtgat tgtcagagca gggaaaatca cctttgcaga aaaggttgca
     1261 aatgctgaaa gcttaaatgc aattggtgtg ttgatataca tggaccagac taaatttccc
     1321 attgttaacg cagaactttc attctttgga catgctcatc tggggacagg tgacccttac
     1381 acacctggat tcccttcctt caatcacact cagtttccac catctcggtc atcaggattg
     1441 cctaatatac ctgtccagac aatctccaga gctgctgcag aaaagctgtt tgggaatatg
     1501 gaaggagact gtccctctga ctggaaaaca gactctacat gtaggatggt aacctcagaa
     1561 agcaagaatg tgaagctcac tgtgagcaat gtgctgaaag agataaaaat tcttaacatc
     1621 tttggagtta ttaaaggctt tgtagaacca gatcactatg ttgtagttgg ggcccagaga
     1681 gatgcatggg gccctggagc tgcaaaatcc ggtgtaggca cagctctcct attgaaactt
     1741 gcccagatgt tctcagatat ggtcttaaaa gatgggtttc agcccagcag aagcattatc
     1801 tttgccagtt ggagtgctgg agactttgga tcggttggtg ccactgaatg gctagaggga
     1861 tacctttcgt ccctgcattt aaaggctttc acttatatta atctggataa agcggttctt
     1921 ggtaccagca acttcaaggt ttctgccagc ccactgttgt atacgcttat tgagaaaaca
     1981 atgcaaaatg tgaagcatcc ggttactggg caatttctat atcaggacag caactgggcc
     2041 agcaaagttg agaaactcac tttagacaat gctgctttcc ctttccttgc atattctgga
     2101 atcccagcag tttctttctg tttttgcgag gacacagatt atccttattt gggtaccacc
     2161 atggacacct ataaggaact gattgagagg attcctgagt tgaacaaagt ggcacgagca
     2221 gctgcagagg tcgctggtca gttcgtgatt aaactaaccc atgatgttga attgaacctg
     2281 gactatgaga ggtacaacag ccaactgctt tcatttgtga gggatctgaa ccaatacaga
     2341 gcagacataa aggaaatggg cctgagttta cagtggctgt attctgctcg tggagacttc
     2401 ttccgtgcta cttccagact aacaacagat ttcgggaatg ctgagaaaac agacagattt
     2461 gtcatgaaga aactcaatga tcgtgtcatg agagtggagt atcacttcct ctctccctac
     2521 gtatctccaa aagagtctcc tttccgacat gtcttctggg gctccggctc tcacacgctg
     2581 ccagctttac tggagaactt gaaactgcgt aaacaaaata acggtgcttt taatgaaacg
     2641 ctgttcagaa accagttggc tctagctact tggactattc agggagctgc aaatgccctc
     2701 tctggtgacg tttgggacat tgacaatgag tttcgggatc caccggtcgc caccatggtg
     2761 tccaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2821 gtaaacggcc acaagttctc cgtgcggggc gagggcgagg gcgatgccac caacggcaag
     2881 ctgaccctga agttcatcag caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2941 accaccctga cctacggcgt gcagagcttc tcccgctacc ccgaccacat gaagcgccac
     3001 gacttcttca agagcgccat gcccgaaggc tacgtccagg agcgcaccat ctccttcaag
     3061 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3121 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3181 gagtacaact tcaactccca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     3241 aaggccaact tcaagatccg ccacaacgtg gaggacggct ccgtgcagct cgccgaccac
     3301 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3361 tccacccagt ccaagctgtc caaagacccc aacgagaagc gcgatcacat ggtccttctg
     3421 gaattcgtga ccgccgccgg gatcactcac ggcatggacg agctgtacaa gtaagcggcc
     3481 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3541 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3601 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3661 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3721 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3781 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3841 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3901 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3961 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     4021 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     4081 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     4141 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     4201 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4261 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     4321 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4381 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4441 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4501 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4561 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4621 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4681 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4741 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4801 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4861 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4921 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4981 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5041 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5101 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5161 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5221 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5281 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5341 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5401 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5461 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5521 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5581 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5641 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5701 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5761 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5821 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5881 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5941 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6001 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6241 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6721 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6781 tgtggataac cgtattaccg cc
//
