LOCUS       sequence-365740-        5927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of N-terminal myc tagged SLC35A2 isoform c/UGT1
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5927)
  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 5927)
  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-myc-SLC35A2
FEATURES             Location/Qualifiers
     source          1..5927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             647..676
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1883..1892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1889..1891,1892..1894,1895..2608)
                     /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(1889..1891,1892..1894,1895..2605)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1934..1955)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2195..2214)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2542..2563
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2731..2852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2768..2787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2822..2841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2859..3314)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2996..3017)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3208..3227
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3341..3445
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3447..3804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3447..3638
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3470..3490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3655..3790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3839..4633
                     /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             3839..4633
                     /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(3893..3912)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4503..4522
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4821..4840)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4865..4912
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5241..5829
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5730..5749
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctgac tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gcgctaccgg actcagatct cgagctcaag cttatggaac aaaaactcat
      661 ctcagaagag gatctgggtg gtggcggctc tatggcagcg gttggggctg gtggttctac
      721 cgctgctcct gggccaggag cggtttccgc gggtgcattg gagccgggaa cagctagtgc
      781 ggctcacagg cgcctgaagt acatatccct agctgtgctg gtggtccaga atgcctccct
      841 catcctcagc atccgctacg cccgcacgtt gccaggggac cgcttctttg ccaccactgc
      901 tgtggtcatg gcggaagtgc tcaaaggcct cacatgcctg ctgctgctct tcgcacagaa
      961 gaggggtaac gtgaagcacc tggttctctt cctccatgag gctgtcctgg tgcagtatgt
     1021 ggacacgctc aagctcgcag tgccctctct catctacacc ttgcagaata acctccagta
     1081 tgttgccatc tctaacctac cagctgccac tttccaggtg acataccagc tgaagatcct
     1141 gaccacagcg ctgttctccg tgctcatgct gaatcgcagt ctttcccggc tgcagtgggc
     1201 ctccctgctg ctcctcttca ctggcgtcgc cattgtccag gcacagcaag ccggtggggg
     1261 aggcccaagg ccactggatc agaaccctgg agcaggcctg gcagctgtcg tggcctcctg
     1321 tctctcctcc ggcttcgcag gagtctactt tgagaagatc ctcaaaggca gctcaggctc
     1381 tgtgtggctg cgcaacctgc aactgggcct cttcggcaca gcactgggcc tggtggggct
     1441 ttggtgggct gagggtaccg ccgttgccac ccgtggtttc ttttttgggt acacacccgc
     1501 tgtctggggc gtggtgctca accaggcctt cggcgggcta ctggtggctg tggttgtcaa
     1561 gtacgctgac aatatcctca agggctttgc cacctccctg tccattgtgc tgtccactgt
     1621 tgcctccatt cgcctctttg gcttccacgt ggacccatta tttgcccttg gcgctggact
     1681 cgtcattggt gctgtctacc tctacagcct tccccgaggt gcagctaaag ccatagcctc
     1741 tgcctctgcc tctgcctccg ggccttgcgt tcaccagcag cctcccgggc agccaccacc
     1801 accgcagctg tcttcccacc gtggagatct catcacggag ccctttctgc caaagtcagt
     1861 gctggtgaag tgaggatcca tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     1921 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1981 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2041 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     2101 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2161 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2221 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2281 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2341 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2401 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2461 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     2521 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2581 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
     2641 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     2701 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     2761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     2821 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     2881 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     2941 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3001 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3061 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3121 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3181 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3241 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3301 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3421 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3481 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3541 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3601 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3661 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3721 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     3781 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     3841 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3901 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3961 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4021 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4081 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4141 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4201 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4261 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4321 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4381 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4441 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4501 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4561 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4621 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4681 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4741 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4801 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4861 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     4921 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4981 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5041 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5101 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5161 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5221 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5281 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5341 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5401 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5461 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5521 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5581 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5641 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5701 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5761 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5821 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5881 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccg
//
