LOCUS       sequence-346747-        7175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression plasmid for a UDP-GlcNAc sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7175)
  AUTHORS   Li Z, Zhang J, Ai HW
  TITLE     Genetically Encoded Green Fluorescent Biosensors for Monitoring 
            UDP-GlcNAc in Live Cells.
  JOURNAL   ACS Cent Sci. 2021 Oct 27;7(10):1763-1770. doi: 
            10.1021/acscentsci.1c00745. Epub 2021 Sep 30.
  PUBMED    34729420
REFERENCE   2  (bases 1 to 7175)
  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: 
            "10.1021/acscentsci.1c00745"; journalName: "ACS Cent Sci"; date: 
            "2021-10-27- 27"; volume: "7"; issue: "10"; pages: "1763-1770"
COMMENT     Sequence Label: pcDNA3-UGAcS
FEATURES             Location/Qualifiers
     source          1..7175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     875..894
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        875..893
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1311..1332
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1444..1465)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2710..2727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2740..2758)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2741..2758)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2778..2795)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2784..3008
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3054..3482
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3141..3160)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3351..3372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3491..3511)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3496..3825
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3676..3811
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3738..3757
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3892..4686
                     /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(3946..3965)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4556..4575
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4860..4981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4897..4916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4951..4970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5030..5046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5030..5046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5043..5065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5054..5070
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5078..5084,5085..5102,5103..5108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5123..5144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5261..5278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5432..6020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5512..5531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6191..6982,6983..7051))
                     /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     6814..6833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7052..7156)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      841 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      901 aagcttgcca ccatgagtgg tcaaggaaag cgattaatgg tgatagcagg cggaaccggt
      961 ggatctgtat tcccgggact ggcggttgcg caccatctaa tggctcaggg ttggcaagtt
     1021 cgctggctgg gaactgccga ccgtatggaa gcggacttag tgccaaaaca tggcatcgaa
     1081 attgatttca ttcgtatctc tggtgaggtt tatatcaagg ccgacaagca gaagaacggc
     1141 atcaaggcga acttccagat ccgccacaac gtggaggacg gcagcatgca gctcgccgac
     1201 cactaccagc agaacacccc cattggcgac ggccccgtgc tgctgcccga caaccattac
     1261 ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1321 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgca caaggtggac
     1381 ggcggatccg gtggcaccgg agtgagcagg ggcgaggagc tgttcaccgg ggtggtgccc
     1441 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tccgcgtgag gggcgaaggc
     1501 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1561 cccgtgccct ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg cttctcccgc
     1621 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1681 caggagcgca ccatcttctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     1741 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1801 ggcaacatcc tggggcacaa gctggagtac gagtctctgc gtggaaaagg tataaaagca
     1861 ctgatagctg ccccgctgcg tatcttcaac gcctgggagc aggcgcgggc gattatgaaa
     1921 gcgtacaaac ctgacgtggt gctcggtatg ggaggctacg tgtcaggtcc aggtggtctg
     1981 gccgcgtggt cgttaggcat tccggttgta cttcatgaac aaaacggtat tgcgggctta
     2041 accaataaat ggctggcgaa gattgccacc aaagtgatgc aggcgtttcc aggtgctttc
     2101 cctaatgcgg aagtagtggg taacccggtg cgtaccgatg tgttggcgct gccgttgccg
     2161 cggcaacgtt tggctggacg tgaaggtccg gttcgtgtgc tggtagtggg tggtacgacg
     2221 ggcgcacgca ttcttaacca gacaatgccg caggttgctg cgaaactggg tgattcagtc
     2281 actatctggc atcagagcgg caaaggttcg caacaatccg ttgaacaggc gtatgccgaa
     2341 gcagggcaac cacagcataa ggtgacggga tttattgatg atatggcggc ggcgtatgcg
     2401 tgggcggatg tcgtcgtttg ccgctccggt gcgttaacgg tgagtgaaat cgccgcggca
     2461 ggactaccgg cgttgtttgt gccgtttcaa cataaagacc gccagcaata ctggaatgcg
     2521 ctaccgctgg aaaaagcggg cgcagccaaa attatcgagc agccacagct tagcgtggat
     2581 gctgtcgcca acaccctggc cgggtggtcg cgagaaacct tattaaccat ggcagaacgc
     2641 gcccgtgctg catccattcc ggatgccacc gagcgagtgg caaatgaagt gagccgggtt
     2701 gcccgggcgc accatcacca ccatcattaa gggccctatt ctatagtgtc acctaaatgc
     2761 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2821 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2881 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2941 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3001 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3061 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3121 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3181 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3241 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3301 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3361 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3421 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3481 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3541 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3661 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3721 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3781 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3841 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3901 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3961 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4021 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4081 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4141 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4201 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4261 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4321 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4381 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4441 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4501 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4561 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4621 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4681 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4741 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4801 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4861 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4921 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4981 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5041 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5101 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5161 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5221 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5281 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5341 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5401 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5461 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5521 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5581 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5641 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5701 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5761 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5821 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5881 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5941 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6001 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6061 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6121 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6181 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6301 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6481 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6541 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6601 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6661 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6721 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6781 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6841 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6901 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6961 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7021 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7081 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7141 aaataggggt tccgcgcaca tttccccgaa aagtg
//
