LOCUS       sequence-365739-        5876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of C-terminal GFP tagged SLC35A2 isoform c/UGT1
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5876)
  AUTHORS   Li D, Mukhopadhyay S
  TITLE     Functional analyses of the UDP-galactose transporter SLC35A2 using 
            the binding of bacterial Shiga toxins as a novel activity assay.
  JOURNAL   Glycobiology. 2019 Jun 1;29(6):490-503. doi: 10.1093/glycob/cwz016.
  PUBMED    30834435
REFERENCE   2  (bases 1 to 5876)
  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"; doi: "10"; journalName: 
            "Glycobiology"; date: "2019-06-1- 1"; volume: "29"; issue: "6"; 
            pages: "490-503"
COMMENT     Sequence Label: pEGFP.N3-SLC35A2-GFP
FEATURES             Location/Qualifiers
     source          1..5876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        89..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        393..596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     547..567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1844..1853
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1850..1852,1853..1855,1856..2569)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1850..1852,1853..1855,1856..2566)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1895..1916)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2156..2175)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2503..2524
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2692..2813
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2729..2748)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2783..2802
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2820..3275)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2957..2978)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3169..3188
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3302..3406
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3408..3765
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3408..3599
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3431..3451)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3616..3751
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3800..4594
                     /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             3800..4594
                     /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(3854..3873)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4464..4483
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4782..4801)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4826..4873
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5202..5790
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5691..5710
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat
       61 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      121 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      421 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      481 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      541 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
      601 tagtgaaccg tcagatccgc tagcgctacc ggactcagat ctcgagctca agcttatggc
      661 agcggttggg gctggtggtt ctaccgctgc tcctgggcca ggagcggttt ccgcgggtgc
      721 attggagccg ggaacagcta gtgcggctca caggcgcctg aagtacatat ccctagctgt
      781 gctggtggtc cagaatgcct ccctcatcct cagcatccgc tacgcccgca cgttgccagg
      841 ggaccgcttc tttgccacca ctgctgtggt catggcggaa gtgctcaaag gcctcacatg
      901 cctgctgctg ctcttcgcac agaagagggg taacgtgaag cacctggttc tcttcctcca
      961 tgaggctgtc ctggtgcagt atgtggacac gctcaagctc gcagtgccct ctctcatcta
     1021 caccttgcag aataacctcc agtatgttgc catctctaac ctaccagctg ccactttcca
     1081 ggtgacatac cagctgaaga tcctgaccac agcgctgttc tccgtgctca tgctgaatcg
     1141 cagtctttcc cggctgcagt gggcctccct gctgctcctc ttcactggcg tcgccattgt
     1201 ccaggcacag caagccggtg ggggaggccc aaggccactg gatcagaacc ctggagcagg
     1261 cctggcagct gtcgtggcct cctgtctctc ctccggcttc gcaggagtct actttgagaa
     1321 gatcctcaaa ggcagctcag gctctgtgtg gctgcgcaac ctgcaactgg gcctcttcgg
     1381 cacagcactg ggcctggtgg ggctttggtg ggctgagggt accgccgttg ccacccgtgg
     1441 tttctttttt gggtacacac ccgctgtctg gggcgtggtg ctcaaccagg ccttcggcgg
     1501 gctactggtg gctgtggttg tcaagtacgc tgacaatatc ctcaagggct ttgccacctc
     1561 cctgtccatt gtgctgtcca ctgttgcctc cattcgcctc tttggcttcc acgtggaccc
     1621 attatttgcc cttggcgctg gactcgtcat tggtgctgtc tacctctaca gccttccccg
     1681 aggtgcagct aaagccatag cctctgcctc tgcctctgcc tccgggcctt gcgttcacca
     1741 gcagcctccc gggcagccac caccaccgca gctgtcttcc caccgtggag atctcatcac
     1801 ggagcccttt ctgccaaagt cagtgctggt gaagggatcc atcgccacca tggtgagcaa
     1861 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1921 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1981 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2041 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2101 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2161 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2221 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2281 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2341 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2401 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2461 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2521 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     2581 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2641 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2701 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2761 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2821 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2881 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2941 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3001 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3061 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3121 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3181 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3241 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3301 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3361 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3421 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3481 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3541 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3601 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3661 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3721 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3781 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3841 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3901 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3961 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4021 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4081 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4141 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4201 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4261 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4321 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4381 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4441 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4501 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4561 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4621 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4681 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4741 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4801 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4861 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4921 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4981 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5041 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5101 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5161 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5221 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5281 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5341 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5401 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5461 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5521 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5581 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5641 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5701 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5761 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5821 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattc
//
