LOCUS       sequence-333152-        9109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for generation of stable mammalian cell lines expressing UAP1 
            with with F383G mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9109)
  AUTHORS   Capota E, Wu H, Kohler JJ
  TITLE     Photocrosslinking O-GlcNAcylated Proteins to Neighboring 
            Biomolecules.
  JOURNAL   Curr Protoc. 2021 Jul;1(7):e201. doi: 10.1002/cpz1.201.
  PUBMED    34288588
REFERENCE   2  (bases 1 to 9109)
  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: "Curr 
            Protoc."; date: "2021-07"; volume: "1(7):e201. doi"; pages: " 
            10.1002/cpz1.201"
COMMENT     Sequence Label: pSin4 UAP1 F383G
FEATURES             Location/Qualifiers
     source          1..9109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    join(2816..3390,3391..3393,3394..3402)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2983..3000)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3210..3229
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3422..4021
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3422..3441)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3918..3938
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4224..4457
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4608..4625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4614..4838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4881..4890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4934..5038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5039..5107,5108..5899)
                     /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     complement(5257..5276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6070..6658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6559..6578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             6817..7450
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7497..7622
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8115..8348
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8533..8577
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8864..8981
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattgatcc ttcgaactag
     1201 tatgaacatt aatgacctca aactcacgtt gtccaaagct gggcaagagc acctactacg
     1261 tttctggaat gagcttgaag aagcccaaca ggtagaactt tatgcagagc tccaggccat
     1321 gaactttgag gagctgaact tctttttcca aaaggccatt gaaggtttta accagtcttc
     1381 tcaccaaaag aatgtggatg cacgaatgga acctgtgcct cgagaggtat taggcagtgc
     1441 tacaagggat caagatcagc tccaggcctg ggaaagtgaa ggacttttcc agatttctca
     1501 gaataaagta gcagttcttc ttctagctgg tgggcagggg acaagactcg gcgttgcata
     1561 tcctaagggg atgtatgatg ttggtttgcc atcccgtaag acactttttc agattcaagc
     1621 agagcgtatc ctgaagctac agcaggttgc tgaaaaatat tatggcaaca aatgcattat
     1681 tccatggtat ataatgacca gtggcagaac aatggaatct acaaaggagt tcttcaccaa
     1741 gcacaagtac tttggtttaa aaaaagagaa tgtaatcttt tttcagcaag gaatgctccc
     1801 cgccatgagt tttgatggga aaattatttt ggaagagaag aacaaagttt ctatggctcc
     1861 agatgggaat ggtggtcttt atcgggcact tgcagcccag aatattgtgg aggatatgga
     1921 gcaaagaggc atttggagca ttcatgtcta ttgtgttgac aacatattag taaaagtggc
     1981 agacccacgg ttcattggat tttgcattca gaaaggagca gactgtggag caaaggtggt
     2041 agagaaaacg aaccctacag aaccagttgg agtggtttgc cgagtggatg gagtttacca
     2101 ggtggtagaa tatagtgaga tttccctggc aacagctcaa aaacgaagct cagacggacg
     2161 actgctgttc aatgcgggga acattgccaa ccatttcttc actgtaccat ttctgagaga
     2221 tgttgtcaat gtttatgaac ctcagttgca gcaccatgtg gctcaaaaga agattcctta
     2281 tgtggatacc caaggacagt taattaagcc agacaaaccc aatggaataa agatggaaaa
     2341 atttgtcggt gacatcttcc agtttgcaaa gaagtttgtg gtatatgaag tattgcgaga
     2401 agatgagttt tccccactaa agaatgctga tagtcagaat gggaaagaca accctactac
     2461 tgcaaggcat gctttgatgt cccttcatca ttgctgggtc ctcaatgcag ggggccattt
     2521 catagatgaa aatggctctc gccttccagc aattccccgc ttgaaggatg ccaatgatgt
     2581 accaatccaa tgtgaaatct ctcctcttat ctcctatgct ggagaaggat tagaaagtta
     2641 tgtggcagat aaagagttcc atgcacctct aatcatcgat gagaatggag ttcatgagct
     2701 ggtgaaaaat ggtatttgag aattcgaagc ttgagctcga gatctgagtc cggtagcgct
     2761 agttcgaagg atccgcatgc atctagggcg gccaattccg cccctctccc ccccccccct
     2821 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2881 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2941 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     3001 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     3061 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     3121 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     3181 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     3241 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     3301 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3361 tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccacaagga gacgaccttc
     3421 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     3481 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     3541 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     3601 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     3661 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     3721 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     3781 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     3841 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     3901 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     3961 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     4021 ataataggcg gccgctcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     4081 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     4141 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     4201 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     4261 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4321 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4381 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4441 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     4501 aaagaaatga atatcagaga gtgagaggcc ttgacattat aatagattta gcaggaattg
     4561 aactaggagt ggagcacaca ggcaaagttc tagagctcgc tgatcagcct cgactgtgcc
     4621 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4681 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4741 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4801 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4861 ctgggggcgc gcccctcgag gccgccatgg tcatagctgt ttgacgtcag gtggcacttt
     4921 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4981 tccgctcacg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5041 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5101 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5161 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5221 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5281 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5341 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5401 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5461 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5521 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5581 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5641 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5701 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5761 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5821 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5881 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6061 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6121 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6181 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6541 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6601 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6661 cgccagcaac gcggcctttt tacggttctg gccttttgca cggaccgcca ctgccaatta
     6721 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
     6781 ttgtatttgt taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa
     6841 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc
     6901 agaactacac accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc
     6961 tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt
     7021 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt
     7081 ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga
     7141 actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg
     7201 gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt
     7261 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     7321 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     7381 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     7441 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     7501 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     7561 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     7621 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     7681 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     7741 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     7801 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     7861 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     7921 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     7981 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     8041 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     8101 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     8161 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     8221 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     8281 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     8341 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     8401 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     8461 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     8521 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     8581 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     8641 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     8701 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     8761 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     8821 ggatctcgac ggtatcgcca caaatggcag tattcatcca caattttaaa agaaaggggg
     8881 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     8941 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     9001 gcagagatcc actttggatc gataagcttt gcaaagatgg ataaagtttt aaacagagag
     9061 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaatt
//
