LOCUS       sequence-365738-        6733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of N-terminal 3×FLAG tagged SLC35A2 isoform 
            c/UGT1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  AUTHORS   Li D, Mukhopadhyay S
  TITLE     A three-pocket model for substrate coordination and selectivity by 
            the nucleotide sugar transporters SLC35A1 and SLC35A2.
  JOURNAL   J Biol Chem. 2021 Sep;297(3):101069. doi: 10.1016/j.jbc.2021.101069.
            Epub 2021 Aug 10.
  PUBMED    34384782
REFERENCE   2  (bases 1 to 6733)
  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 Biol 
            Chem."; date: "2021-09"; volume: "297(3):101069. doi"; pages: " 
            10.1016/j.jbc.2021.101069. Epub 2021 Aug 10"
COMMENT     Sequence Label: pCMV-3×FLAG-SLC35A2
FEATURES             Location/Qualifiers
     source          1..6733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..178
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     170..187
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     171..187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        348..727
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        728..931
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     882..902
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             961..1026
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2219..2841
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2418..2437)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2865..2885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2883..3180
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3234..4028
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3288..3307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3898..3917
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4557..5145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4637..4656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5316..6107,6108..6176))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5939..5958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6177..6281)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6308..6733,1..3)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6395..6414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6605..6626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg
       61 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
      121 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
      181 ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat
      241 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
      301 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt
      361 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      421 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      481 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      541 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      601 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      661 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      721 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
      781 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      841 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
      901 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga attaaccatg
      961 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     1021 gacaagctta tggcagcggt tggggctggt ggttctaccg ctgctcctgg gccaggagcg
     1081 gtttccgcgg gtgcattgga gccgggaaca gctagtgcgg ctcacaggcg cctgaagtac
     1141 atatccctag ctgtgctggt ggtccagaat gcctccctca tcctcagcat ccgctacgcc
     1201 cgcacgttgc caggggaccg cttctttgcc accactgctg tggtcatggc ggaagtgctc
     1261 aaaggcctca catgcctgct gctgctcttc gcacagaaga ggggtaacgt gaagcacctg
     1321 gttctcttcc tccatgaggc tgtcctggtg cagtatgtgg acacgctcaa gctcgcagtg
     1381 ccctctctca tctacacctt gcagaataac ctccagtatg ttgccatctc taacctacca
     1441 gctgccactt tccaggtgac ataccagctg aagatcctga ccacagcgct gttctccgtg
     1501 ctcatgctga atcgcagtct ttcccggctg cagtgggcct ccctgctgct cctcttcact
     1561 ggcgtcgcca ttgtccaggc acagcaagcc ggtgggggag gcccaaggcc actggatcag
     1621 aaccctggag caggcctggc agctgtcgtg gcctcctgtc tctcctccgg cttcgcagga
     1681 gtctactttg agaagatcct caaaggcagc tcaggctctg tgtggctgcg caacctgcaa
     1741 ctgggcctct tcggcacagc actgggcctg gtggggcttt ggtgggctga gggtaccgcc
     1801 gttgccaccc gtggtttctt ttttgggtac acacccgctg tctggggcgt ggtgctcaac
     1861 caggccttcg gcgggctact ggtggctgtg gttgtcaagt acgctgacaa tatcctcaag
     1921 ggctttgcca cctccctgtc cattgtgctg tccactgttg cctccattcg cctctttggc
     1981 ttccacgtgg acccattatt tgcccttggc gctggactcg tcattggtgc tgtctacctc
     2041 tacagccttc cccgaggtgc agctaaagcc atagcctctg cctctgcctc tgcctccggg
     2101 ccttgcgttc accagcagcc tcccgggcag ccaccaccac cgcagctgtc ttcccaccgt
     2161 ggagatctca tcacggagcc ctttctgcca aagtcagtgc tggtgaagtg aggatcccgg
     2221 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2281 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2341 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2401 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2461 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2521 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2581 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     2641 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     2701 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     2761 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     2821 tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt
     2881 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2941 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3001 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3061 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3121 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3181 aggaattgat cagcttggga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3241 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3301 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3361 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3421 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3481 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3541 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3601 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3661 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3721 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3781 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3841 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3901 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3961 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4021 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     4081 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     4141 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     4201 cgggctcgat cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga
     4261 gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat
     4321 ccatgccccc gaactgcagg agtggggagg cacgatggcc gctttggtcg acccggacgg
     4381 gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc
     4441 cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag
     4501 ctgctcacca cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4741 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4981 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6301 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6361 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6421 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6481 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6541 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6601 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6661 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6721 aaatttaacg cga
//
