LOCUS       sequence-398164-        7908 bp    DNA     circular SYN 14-APR-2025
DEFINITION  boronic acid- and peptide-assisted UDP-GlcNAc sensor in ER.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7908)
  AUTHORS   Zhang J, Li Z, Pang Y, Fan Y, Ai HW
  TITLE     Genetically Encoded Boronolectin as a Specific Red Fluorescent 
            UDP-GlcNAc Biosensor.
  JOURNAL   ACS Sens. 2023 Jul 22. doi: 10.1021/acssensors.3c00409.
  PUBMED    37480329
REFERENCE   2  (bases 1 to 7908)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Sens. 
            2023 Jul 22. doi: 10.1021/acssensors.3c00409."
COMMENT     Sequence Label: pMAH-ER-bapaUGAc
FEATURES             Location/Qualifiers
     source          1..7908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        739..757
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     739..756
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     770..789
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1639..1658
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1790..1807)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1796..2020
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2066..2494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2153..2172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2363..2384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2503..2523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2508..2837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2688..2823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2750..2769
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2886..3911
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2886..3911
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4041..4162
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4078..4097)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4132..4151
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4211..4227)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4211..4227)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4224..4246)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4235..4251
                     /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(4259..4265,4266..4283,4284..4289))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4304..4325
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4442..4459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4613..5201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4693..4712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5372..6163,6164..6232))
                     /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     5995..6014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6233..6337)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(6461..6551)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(6663..6911)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(7200..7290)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(7402..7650)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccgcggccg
      601 ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta ccgcctatag
      661 agtctatagg cccaccccct tggcttcgtt agaacgcggc tacaattaat acataacctt
      721 atgtatcata cacatacgat ttaggtgaca ctatagaata acatccactt tgcctttctc
      781 tccacaggtg tccactccca ggtccaactg cacggaagct tgccccatgc tgctatccgt
      841 gccgctgctg ctcggcctgc tgggcctggc cgcagctgac gaattcgatt cttattttgg
      901 aaaaacttat aatccatggg atcctatgca gcggaccgtg tacatctccg agtccgagcg
      961 gatgtacccc gaggacggcg ccctgaagag cgagatcaag taggggctga ggctgaagga
     1021 cggcggccac tacgccgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     1081 gcccggcgcc tacatcgtcg acatcaagtt ggacatcgtg tcccacaacg aggactacac
     1141 catcgtggaa cagtgcgaac gcgccgaggg ccgccactcc actggcggca tggacgagct
     1201 gtacaaggga ggtacaggcg ggagtctggt gagcaagggc gaggaggata acatggccat
     1261 catcaaggag ttcatgagct tcaaggtgca catggagggc tccgtgaacg gccacaagtt
     1321 cgagatcgag ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa
     1381 ggtgaccatc ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     1441 cggctccaag gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt
     1501 ccccgagggc ttcaggtggg agcgcgtgat gaacttcgag gacggcggca ttattcacgt
     1561 taaccaggac tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac
     1621 caacttcccc cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggctacgcc
     1681 tacgcctcgg gtggtgggtg atttatataa gaatggtatt gatccatctt atactcctca
     1741 tcgtgaccaa aaggacgaac tctagggccc gtttaaaccc gctgatcagc ctcgactgtg
     1801 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1861 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1921 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1981 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2041 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2101 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2161 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2221 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2281 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2341 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2401 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2461 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2521 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2581 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2641 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     2701 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     2761 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     2821 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     2881 acgtgatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     2941 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     3001 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     3061 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     3121 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     3181 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     3241 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     3301 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     3361 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     3421 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     3481 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     3541 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     3601 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     3661 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     3721 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     3781 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     3841 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     3901 caaaggaata gcacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     3961 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4021 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4081 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4141 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4201 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4261 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4321 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4381 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4441 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4501 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4561 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4621 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4681 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4741 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4801 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4861 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4921 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4981 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5041 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5101 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5161 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5221 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5281 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5341 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5401 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5461 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5521 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5581 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5641 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5701 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5761 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5821 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5881 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5941 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6001 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6061 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6121 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6181 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6241 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6301 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga ctcgactcgc
     6361 tagtgttcca aaaaatggtg ggggacggat tcgaaccgcc gaacccaaag ggagcggatt
     6421 tagagtccgc ggcgtttggc cacttcgcta ccccaccaga gtggtctcat acagaactta
     6481 taagattccc aaatccaaag acatttcacg tttatggtga tttcccagaa cacatagcga
     6541 catgcaaata tgaattgatg actgcctgat ggagaaaaaa tggtggggga cggattcgaa
     6601 ccgccgaacc caaagggagc ggatttagag tccgcggcgt ttggccactt cgctacccca
     6661 ccggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt
     6721 acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga
     6781 aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta
     6841 attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga
     6901 aataggccct cttcctgccc gaccttcgcg atgtacgggc cagatatacg ccaatattgg
     6961 ccattagcca tattattcat tggttatata gcataaatca atattggcta ttggccattg
     7021 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
     7081 ccatgttggc tcgactcgct agtgttccaa aaaatggtgg gggacggatt cgaaccgccg
     7141 aacccaaagg gagcggattt agagtccgcg gcgtttggcc acttcgctac cccaccagag
     7201 tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat
     7261 ttcccagaac acatagcgac atgcaaatat gaattgatga ctgcctgatg gagaaaaaat
     7321 ggtgggggac ggattcgaac cgccgaaccc aaagggagcg gatttagagt ccgcggcgtt
     7381 tggccacttc gctaccccac cggtgtttcg tcctttccac aagatatata aagccaagaa
     7441 atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac ataattttaa
     7501 aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt
     7561 tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc gtatatgcaa
     7621 atatgaagga atcatgggaa ataggccctc ttcctgcccg accttcgcga tgtacgggcc
     7681 agatatacgc caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     7741 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     7801 ctcatgtcca atatgaccgc catgttggct cgagcattga ttattgacta gttattaata
     7861 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
