LOCUS       sequence-365737-        6565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of N-terminal 3×FLAG tagged SLC35A1 isoform a 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  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 6565)
  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-SLC35A1
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..153
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     145..162
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     146..162
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        323..702
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        703..906
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     857..877
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             936..1001
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2026..2648
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2225..2244)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2672..2692)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2690..2987
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3041..3835
                     /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(3095..3114)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3705..3724
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4364..4952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4444..4463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5123..5914,5915..5983))
                     /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     5746..5765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5984..6088)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6115..6543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6202..6221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6412..6433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
       61 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      121 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct gatctataca
      181 ttgaatcaat attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt
      241 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
      301 tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
      361 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      421 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      481 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      541 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
      601 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
      661 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      721 tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      781 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat
      841 aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      901 agagctcgtt tagtgaaccg tcagaattaa ccatggacta caaagaccat gacggtgatt
      961 ataaagatca tgacatcgat tacaaggatg acgatgacaa gcttatggct gccccgagag
     1021 acaatgtcac tttattattc aagttatact gcttggcagt gatgaccctg atggctgcag
     1081 tctataccat agctttaaga tacacaagga catcagacaa agaactctac ttttcaacca
     1141 cagccgtgtg tatcacagaa gttataaagt tattgctaag tgtgggaatt ttagctaaag
     1201 aaactggtag tctgggtaga ttcaaagcat ctttaagaga aaatgtcttg gggagcccca
     1261 aggaactgtt gaagttaagt gtgccatcgt tagtgtatgc tgttcagaac aacatggctt
     1321 tcctagctct tagcaatctg gatgcagcag tgtaccaggt gacctaccag ttgaagattc
     1381 cgtgtactgc tttatgcact gttttaatgt taaaccggac actcagcaaa ttacagtggg
     1441 tttcagtttt tatgctgtgt gctggagtta cgcttgtaca gtggaaacca gcccaagcta
     1501 caaaagtggt ggtggaacaa aatccattat tagggtttgg cgctatagct attgctgtat
     1561 tgtgctcagg atttgcagga gtatattttg aaaaagtttt aaagagttca gatacttctc
     1621 tttgggtgag aaacattcaa atgtatctat cagggattat tgtgacatta gctggcgtct
     1681 acttgtcaga tggagctgaa attaaagaaa aaggattttt ctatggttac acatattatg
     1741 tctggtttgt catctttctt gcaagtgttg gtggcctcta cacttctgtt gtggttaagt
     1801 acacagacaa catcatgaaa ggcttttctg cagcagcggc cattgtcctt tccaccattg
     1861 cttcagtaat gctgtttgga ttacagataa cactcacctt tgccctgggt actcttcttg
     1921 tatgtgtttc catatatctc tatggattac ccagacaaga cactacatcc atccaacaag
     1981 gagaaacagc ttcaaaggag agagttattg gtgtgtgagg atcccgggtg gcatccctgt
     2041 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     2101 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     2161 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     2221 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     2281 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     2341 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     2401 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     2461 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta
     2521 taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga
     2581 catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct
     2641 gttgaattgg gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag
     2701 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2761 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2821 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2881 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2941 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg aattgatcag
     3001 cttgggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3061 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3121 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3181 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3241 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3301 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3361 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3421 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3481 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3541 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3601 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3661 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3721 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3781 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3841 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3901 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3961 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccgg gctcgatccc
     4021 ctcgcgagtt ggttcagctg ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag
     4081 tgcaaatccg tcggcatcca ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa
     4141 ctgcaggagt ggggaggcac gatggccgct ttggtcgacc cggacgggac gctcctgcgc
     4201 ctgatacaga acgaattgct tgcaggcatc tcatgagtgt gtcttcccgt tttccgcctg
     4261 aggtcactgc gtggatggag cgctggcgcc tgctgcgcga cggcgagctg ctcaccaccc
     4321 actcgccaag ctggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4801 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4921 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4981 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5041 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5101 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5161 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5221 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5281 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5341 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5401 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5461 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5521 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5581 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5641 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5701 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5761 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5821 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5881 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5941 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6001 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6061 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc
     6121 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6181 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6241 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6301 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6361 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6421 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6481 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6541 attttaacaa aatattaacg cttac
//
