LOCUS       sequence-322790-        6103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with the cytosolic tail 
            and 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 6103)
  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 6103)
  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: FFS-GFP
FEATURES             Location/Qualifiers
     source          1..6103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2068..2077
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2074..2076,2077..2079,2080..2793)
                     /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(2119..2140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2915..3036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2952..2971)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3006..3025
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3043..3498)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3180..3201)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3392..3411
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3525..3629
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3631..3988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3631..3822
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3654..3674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3839..3974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4023..4817
                     /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             4023..4817
                     /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(4077..4096)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4687..4706
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5005..5024)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5049..5096
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5425..6013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5914..5933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      601 gaaccgtcag atccgctagc gctaccggac tcagatctcg aggccaccat gatggatcaa
      661 gctagatcag cattctctaa cttgtttggt ggagaaccat tgtcatatac ccggttcagc
      721 ctggctcggc aagtagatgg cgataacagt catgtggaga tgaaacttgc tgtagatgaa
      781 gaagaaaatg ctgacaataa cacaaaggcc aatgtcacaa aaccaaaaag gtgtagtgga
      841 agtatctgct atgggactat tgctgtgatc gtctttttct tgattggatt tatgattggc
      901 tacttgggct ataaggaaaa gaagaaaggg agttactatg attcctttaa attgcaaacc
      961 aaggaattcc aggtgttaaa gagtctgggg aaattggcca tggggtctga ttcccagtct
     1021 gtatcctcaa gcagcaccca ggacccccac aggggccgcc agaccctcgg cagtctcaga
     1081 ggcctagcca aggccaaacc agaggcctcc ttccaggtgt ggaacaagga cagctcttcc
     1141 aaaaacctta tccctaggct gcaaaagatc tggaagaatt acctaagcat gaacaagtac
     1201 aaagtgtcct acaaggggcc aggaccaggc atcaagttca gtgcagaggc cctgcgctgc
     1261 cacctccggg accatgtgaa tgtatccatg gtagaggtca cagattttcc cttcaatacc
     1321 tctgaatggg agggttatct gcccaaggag agcattagga ccaaggctgg gccttggggc
     1381 aggtgtgctg ttgtgtcgtc agcgggatct ctgaagtcct cccaactagg cagagaaatc
     1441 gatgatcatg acgcagtcct gaggtttaat ggggcaccca cagccaactt ccaacaagat
     1501 gtgggcacaa aaactaccat tcgcctgatg aactctcagt tggttaccac agagaagcgc
     1561 ttcctcaaag acagtttgta caatgaagga atcctaattg tatgggaccc atctgtatac
     1621 cactcagata tcccaaagtg gtaccagaat ccggattata atttctttaa caactacaag
     1681 acttatcgta agctgcaccc caatcagccc ttttacatcc tcaagcccca gatgccttgg
     1741 gagctatggg acattcttca agaaatctcc ccagaagaga ttcagccaaa ccccccatcc
     1801 tctgggatgc ttggtatcat catcatgatg acgctgtgtg accaggtgga tatttatgag
     1861 ttcctcccat ccaagcgcaa gactgacgtg tgctactact accagaagtt cttcgatagt
     1921 gcctgcacga tgggtgccta ccacccgctg ctctatgaga agaatttggt gaagcatctc
     1981 aaccagggca cagatgagga catctacctg cttggaaaag ccacactgcc tggcttccgg
     2041 accattcact gccgggatcc accggtcgcc accatggtgt ccaagggcga ggagctgttc
     2101 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttctcc
     2161 gtgcggggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatcagc
     2221 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2281 cagagcttct cccgctaccc cgaccacatg aagcgccacg acttcttcaa gagcgccatg
     2341 cccgaaggct acgtccagga gcgcaccatc tccttcaagg acgacggcac ctacaagacc
     2401 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2461 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt caactcccac
     2521 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     2581 cacaacgtgg aggacggctc cgtgcagctc gccgaccact accagcagaa cacccccatc
     2641 ggcgacggcc ccgtgctgct gcccgacaac cactacctgt ccacccagtc caagctgtcc
     2701 aaagacccca acgagaagcg cgatcacatg gtccttctgg aattcgtgac cgccgccggg
     2761 atcactcacg gcatggacga gctgtacaag taagcggccg cgactctaga tcataatcag
     2821 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2881 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2941 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3001 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3061 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3121 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3181 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3241 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3301 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3361 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3421 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3481 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3541 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3601 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3661 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3721 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3781 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3841 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3901 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3961 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4021 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4081 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4141 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4201 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4261 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4321 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4381 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4441 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4501 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4561 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4621 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4681 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4741 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4801 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4861 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4921 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4981 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5041 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5101 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5161 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5221 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5281 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5341 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtg
//
